CN109609519A - 一种基因RKcrtYB及其应用 - Google Patents
一种基因RKcrtYB及其应用 Download PDFInfo
- Publication number
- CN109609519A CN109609519A CN201811281423.XA CN201811281423A CN109609519A CN 109609519 A CN109609519 A CN 109609519A CN 201811281423 A CN201811281423 A CN 201811281423A CN 109609519 A CN109609519 A CN 109609519A
- Authority
- CN
- China
- Prior art keywords
- leu
- ala
- gene
- arg
- rkcrtyb
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 28
- 238000004519 manufacturing process Methods 0.000 claims abstract description 8
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 239000002773 nucleotide Substances 0.000 claims abstract description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract 2
- 238000003259 recombinant expression Methods 0.000 claims description 4
- 241000221523 Rhodotorula toruloides Species 0.000 abstract description 24
- 235000021466 carotenoid Nutrition 0.000 abstract description 19
- 150000001747 carotenoids Chemical class 0.000 abstract description 19
- 241000007100 Rhodotorula kratochvilovae Species 0.000 abstract description 7
- 230000015572 biosynthetic process Effects 0.000 abstract description 7
- 238000003786 synthesis reaction Methods 0.000 abstract description 6
- 239000013612 plasmid Substances 0.000 description 16
- 108020004414 DNA Proteins 0.000 description 10
- 230000001580 bacterial effect Effects 0.000 description 10
- 230000029087 digestion Effects 0.000 description 10
- 238000012408 PCR amplification Methods 0.000 description 9
- OENHQHLEOONYIE-UKMVMLAPSA-N all-trans beta-carotene Natural products CC=1CCCC(C)(C)C=1/C=C/C(/C)=C/C=C/C(/C)=C/C=C/C=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C OENHQHLEOONYIE-UKMVMLAPSA-N 0.000 description 6
- 235000013734 beta-carotene Nutrition 0.000 description 6
- TUPZEYHYWIEDIH-WAIFQNFQSA-N beta-carotene Natural products CC(=C/C=C/C=C(C)/C=C/C=C(C)/C=C/C1=C(C)CCCC1(C)C)C=CC=C(/C)C=CC2=CCCCC2(C)C TUPZEYHYWIEDIH-WAIFQNFQSA-N 0.000 description 6
- 239000011648 beta-carotene Substances 0.000 description 6
- 229960002747 betacarotene Drugs 0.000 description 6
- OENHQHLEOONYIE-JLTXGRSLSA-N β-Carotene Chemical compound CC=1CCCC(C)(C)C=1\C=C\C(\C)=C\C=C\C(\C)=C\C=C\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C OENHQHLEOONYIE-JLTXGRSLSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 239000012634 fragment Substances 0.000 description 5
- 108010050848 glycylleucine Proteins 0.000 description 5
- 230000009182 swimming Effects 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 150000001413 amino acids Chemical group 0.000 description 4
- 230000003078 antioxidant effect Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 238000004925 denaturation Methods 0.000 description 4
- 230000036425 denaturation Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 230000002018 overexpression Effects 0.000 description 4
- 108091008146 restriction endonucleases Proteins 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 3
- 241000880493 Leptailurus serval Species 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 235000006708 antioxidants Nutrition 0.000 description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000000284 extract Substances 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 238000003208 gene overexpression Methods 0.000 description 3
- 239000003550 marker Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 229910052760 oxygen Inorganic materials 0.000 description 3
- 239000001301 oxygen Substances 0.000 description 3
- 150000003254 radicals Chemical class 0.000 description 3
- 230000009261 transgenic effect Effects 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 2
- KTXKIYXZQFWJKB-VZFHVOOUSA-N Ala-Thr-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O KTXKIYXZQFWJKB-VZFHVOOUSA-N 0.000 description 2
- KRQSPVKUISQQFS-FJXKBIBVSA-N Arg-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCN=C(N)N KRQSPVKUISQQFS-FJXKBIBVSA-N 0.000 description 2
- MEFGKQUUYZOLHM-GMOBBJLQSA-N Asn-Arg-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MEFGKQUUYZOLHM-GMOBBJLQSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- GRRNUXAQVGOGFE-UHFFFAOYSA-N Hygromycin-B Natural products OC1C(NC)CC(N)C(O)C1OC1C2OC3(C(C(O)C(O)C(C(N)CO)O3)O)OC2C(O)C(CO)O1 GRRNUXAQVGOGFE-UHFFFAOYSA-N 0.000 description 2
- RRHGJUQNOFWUDK-UHFFFAOYSA-N Isoprene Chemical compound CC(=C)C=C RRHGJUQNOFWUDK-UHFFFAOYSA-N 0.000 description 2
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 2
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- ONORAGIFHNAADN-LLLHUVSDSA-N Phe-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N ONORAGIFHNAADN-LLLHUVSDSA-N 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000004087 circulation Effects 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 239000013613 expression plasmid Substances 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 2
- 108010049041 glutamylalanine Proteins 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- GRRNUXAQVGOGFE-NZSRVPFOSA-N hygromycin B Chemical compound O[C@@H]1[C@@H](NC)C[C@@H](N)[C@H](O)[C@H]1O[C@H]1[C@H]2O[C@@]3([C@@H]([C@@H](O)[C@@H](O)[C@@H](C(N)CO)O3)O)O[C@H]2[C@@H](O)[C@@H](CO)O1 GRRNUXAQVGOGFE-NZSRVPFOSA-N 0.000 description 2
- 229940097277 hygromycin b Drugs 0.000 description 2
- 230000003834 intracellular effect Effects 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000000243 photosynthetic effect Effects 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 108010031719 prolyl-serine Proteins 0.000 description 2
- 210000002966 serum Anatomy 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- 239000013598 vector Substances 0.000 description 2
- AUXMWYRZQPIXCC-KNIFDHDWSA-N (2s)-2-amino-4-methylpentanoic acid;(2s)-2-aminopropanoic acid Chemical compound C[C@H](N)C(O)=O.CC(C)C[C@H](N)C(O)=O AUXMWYRZQPIXCC-KNIFDHDWSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 241000589158 Agrobacterium Species 0.000 description 1
- UWQJHXKARZWDIJ-ZLUOBGJFSA-N Ala-Ala-Cys Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CS)C(O)=O UWQJHXKARZWDIJ-ZLUOBGJFSA-N 0.000 description 1
- SSSROGPPPVTHLX-FXQIFTODSA-N Ala-Arg-Asp Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O SSSROGPPPVTHLX-FXQIFTODSA-N 0.000 description 1
- JBGSZRYCXBPWGX-BQBZGAKWSA-N Ala-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N JBGSZRYCXBPWGX-BQBZGAKWSA-N 0.000 description 1
- NWVVKQZOVSTDBQ-CIUDSAMLSA-N Ala-Glu-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NWVVKQZOVSTDBQ-CIUDSAMLSA-N 0.000 description 1
- PAIHPOGPJVUFJY-WDSKDSINSA-N Ala-Glu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O PAIHPOGPJVUFJY-WDSKDSINSA-N 0.000 description 1
- BEMGNWZECGIJOI-WDSKDSINSA-N Ala-Gly-Glu Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O BEMGNWZECGIJOI-WDSKDSINSA-N 0.000 description 1
- QHASENCZLDHBGX-ONGXEEELSA-N Ala-Gly-Phe Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QHASENCZLDHBGX-ONGXEEELSA-N 0.000 description 1
- NBTGEURICRTMGL-WHFBIAKZSA-N Ala-Gly-Ser Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O NBTGEURICRTMGL-WHFBIAKZSA-N 0.000 description 1
- SUMYEVXWCAYLLJ-GUBZILKMSA-N Ala-Leu-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O SUMYEVXWCAYLLJ-GUBZILKMSA-N 0.000 description 1
- PMQXMXAASGFUDX-SRVKXCTJSA-N Ala-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@H](C)N)CCCCN PMQXMXAASGFUDX-SRVKXCTJSA-N 0.000 description 1
- DEWWPUNXRNGMQN-LPEHRKFASA-N Ala-Met-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N1CCC[C@@H]1C(=O)O)N DEWWPUNXRNGMQN-LPEHRKFASA-N 0.000 description 1
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 1
- WQKAQKZRDIZYNV-VZFHVOOUSA-N Ala-Ser-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WQKAQKZRDIZYNV-VZFHVOOUSA-N 0.000 description 1
- CLOMBHBBUKAUBP-LSJOCFKGSA-N Ala-Val-His Chemical compound C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N CLOMBHBBUKAUBP-LSJOCFKGSA-N 0.000 description 1
- PEFFAAKJGBZBKL-NAKRPEOUSA-N Arg-Ala-Ile Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O PEFFAAKJGBZBKL-NAKRPEOUSA-N 0.000 description 1
- OLDOLPWZEMHNIA-PJODQICGSA-N Arg-Ala-Trp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O OLDOLPWZEMHNIA-PJODQICGSA-N 0.000 description 1
- UXJCMQFPDWCHKX-DCAQKATOSA-N Arg-Arg-Glu Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O UXJCMQFPDWCHKX-DCAQKATOSA-N 0.000 description 1
- DJAIOAKQIOGULM-DCAQKATOSA-N Arg-Glu-Met Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O DJAIOAKQIOGULM-DCAQKATOSA-N 0.000 description 1
- LRPZJPMQGKGHSG-XGEHTFHBSA-N Arg-Ser-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)N)O LRPZJPMQGKGHSG-XGEHTFHBSA-N 0.000 description 1
- KEZVOBAKAXHMOF-GUBZILKMSA-N Arg-Val-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N KEZVOBAKAXHMOF-GUBZILKMSA-N 0.000 description 1
- SUMJNGAMIQSNGX-TUAOUCFPSA-N Arg-Val-Pro Chemical compound CC(C)[C@H](NC(=O)[C@@H](N)CCCNC(N)=N)C(=O)N1CCC[C@@H]1C(O)=O SUMJNGAMIQSNGX-TUAOUCFPSA-N 0.000 description 1
- NLCDVZJDEXIDDL-BIIVOSGPSA-N Asn-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC(=O)N)N)C(=O)O NLCDVZJDEXIDDL-BIIVOSGPSA-N 0.000 description 1
- OLISTMZJGQUOGS-GMOBBJLQSA-N Asn-Ile-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N OLISTMZJGQUOGS-GMOBBJLQSA-N 0.000 description 1
- CMCIMCAQIULNDJ-CIUDSAMLSA-N Asp-His-Cys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)O)N CMCIMCAQIULNDJ-CIUDSAMLSA-N 0.000 description 1
- GBSUGIXJAAKZOW-GMOBBJLQSA-N Asp-Ile-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O GBSUGIXJAAKZOW-GMOBBJLQSA-N 0.000 description 1
- UJGRZQYSNYTCAX-SRVKXCTJSA-N Asp-Leu-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC(O)=O UJGRZQYSNYTCAX-SRVKXCTJSA-N 0.000 description 1
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 1
- XYPJXLLXNSAWHZ-SRVKXCTJSA-N Asp-Ser-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O XYPJXLLXNSAWHZ-SRVKXCTJSA-N 0.000 description 1
- PDIYGFYAMZZFCW-JIOCBJNQSA-N Asp-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N)O PDIYGFYAMZZFCW-JIOCBJNQSA-N 0.000 description 1
- JGLWFWXGOINXEA-YDHLFZDLSA-N Asp-Val-Tyr Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 JGLWFWXGOINXEA-YDHLFZDLSA-N 0.000 description 1
- 208000002177 Cataract Diseases 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 235000004237 Crocus Nutrition 0.000 description 1
- 241000596148 Crocus Species 0.000 description 1
- VNLYIYOYUNGURO-ZLUOBGJFSA-N Cys-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CS)N VNLYIYOYUNGURO-ZLUOBGJFSA-N 0.000 description 1
- 102000012410 DNA Ligases Human genes 0.000 description 1
- 108010061982 DNA Ligases Proteins 0.000 description 1
- 108010042407 Endonucleases Proteins 0.000 description 1
- 102000004533 Endonucleases Human genes 0.000 description 1
- 108020004460 Fungal RNA Proteins 0.000 description 1
- 108091092584 GDNA Proteins 0.000 description 1
- IOFDDSNZJDIGPB-GVXVVHGQSA-N Gln-Leu-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O IOFDDSNZJDIGPB-GVXVVHGQSA-N 0.000 description 1
- XKPACHRGOWQHFH-IRIUXVKKSA-N Gln-Thr-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O XKPACHRGOWQHFH-IRIUXVKKSA-N 0.000 description 1
- RUFHOVYUYSNDNY-ACZMJKKPSA-N Glu-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(O)=O RUFHOVYUYSNDNY-ACZMJKKPSA-N 0.000 description 1
- AVZHGSCDKIQZPQ-CIUDSAMLSA-N Glu-Arg-Ala Chemical compound C[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCC(O)=O)C(O)=O AVZHGSCDKIQZPQ-CIUDSAMLSA-N 0.000 description 1
- PXHABOCPJVTGEK-BQBZGAKWSA-N Glu-Gln-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O PXHABOCPJVTGEK-BQBZGAKWSA-N 0.000 description 1
- CGOHAEBMDSEKFB-FXQIFTODSA-N Glu-Glu-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O CGOHAEBMDSEKFB-FXQIFTODSA-N 0.000 description 1
- HNVFSTLPVJWIDV-CIUDSAMLSA-N Glu-Glu-Gln Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O HNVFSTLPVJWIDV-CIUDSAMLSA-N 0.000 description 1
- BDISFWMLMNBTGP-NUMRIWBASA-N Glu-Thr-Asp Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O BDISFWMLMNBTGP-NUMRIWBASA-N 0.000 description 1
- GPSHCSTUYOQPAI-JHEQGTHGSA-N Glu-Thr-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O GPSHCSTUYOQPAI-JHEQGTHGSA-N 0.000 description 1
- RJIVPOXLQFJRTG-LURJTMIESA-N Gly-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N RJIVPOXLQFJRTG-LURJTMIESA-N 0.000 description 1
- CQZDZKRHFWJXDF-WDSKDSINSA-N Gly-Gln-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CN CQZDZKRHFWJXDF-WDSKDSINSA-N 0.000 description 1
- SOEATRRYCIPEHA-BQBZGAKWSA-N Gly-Glu-Glu Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O SOEATRRYCIPEHA-BQBZGAKWSA-N 0.000 description 1
- XTQFHTHIAKKCTM-YFKPBYRVSA-N Gly-Glu-Gly Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O XTQFHTHIAKKCTM-YFKPBYRVSA-N 0.000 description 1
- SXJHOPPTOJACOA-QXEWZRGKSA-N Gly-Ile-Arg Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CCCN=C(N)N SXJHOPPTOJACOA-QXEWZRGKSA-N 0.000 description 1
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 1
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 1
- IZVICCORZOSGPT-JSGCOSHPSA-N Gly-Val-Tyr Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IZVICCORZOSGPT-JSGCOSHPSA-N 0.000 description 1
- AWHJQEYGWRKPHE-LSJOCFKGSA-N His-Ala-Arg Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AWHJQEYGWRKPHE-LSJOCFKGSA-N 0.000 description 1
- SKYULSWNBYAQMG-IHRRRGAJSA-N His-Leu-Arg Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SKYULSWNBYAQMG-IHRRRGAJSA-N 0.000 description 1
- OWYIDJCNRWRSJY-QTKMDUPCSA-N His-Pro-Thr Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O OWYIDJCNRWRSJY-QTKMDUPCSA-N 0.000 description 1
- UOYGZBIPZYKGSH-SRVKXCTJSA-N His-Ser-Lys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)O)N UOYGZBIPZYKGSH-SRVKXCTJSA-N 0.000 description 1
- TZCGZYWNIDZZMR-UHFFFAOYSA-N Ile-Arg-Ala Natural products CCC(C)C(N)C(=O)NC(C(=O)NC(C)C(O)=O)CCCN=C(N)N TZCGZYWNIDZZMR-UHFFFAOYSA-N 0.000 description 1
- HPCFRQWLTRDGHT-AJNGGQMLSA-N Ile-Leu-Leu Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O HPCFRQWLTRDGHT-AJNGGQMLSA-N 0.000 description 1
- CIDLJWVDMNDKPT-FIRPJDEBSA-N Ile-Phe-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)O)N CIDLJWVDMNDKPT-FIRPJDEBSA-N 0.000 description 1
- CAHCWMVNBZJVAW-NAKRPEOUSA-N Ile-Pro-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)O)N CAHCWMVNBZJVAW-NAKRPEOUSA-N 0.000 description 1
- AKQFLPNANHNTLP-VKOGCVSHSA-N Ile-Pro-Trp Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O)N AKQFLPNANHNTLP-VKOGCVSHSA-N 0.000 description 1
- WCNWGAUZWWSYDG-SVSWQMSJSA-N Ile-Thr-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)O)N WCNWGAUZWWSYDG-SVSWQMSJSA-N 0.000 description 1
- SITWEMZOJNKJCH-UHFFFAOYSA-N L-alanine-L-arginine Natural products CC(N)C(=O)NC(C(O)=O)CCCNC(N)=N SITWEMZOJNKJCH-UHFFFAOYSA-N 0.000 description 1
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 1
- QPRQGENIBFLVEB-BJDJZHNGSA-N Leu-Ala-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O QPRQGENIBFLVEB-BJDJZHNGSA-N 0.000 description 1
- XIRYQRLFHWWWTC-QEJZJMRPSA-N Leu-Ala-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 XIRYQRLFHWWWTC-QEJZJMRPSA-N 0.000 description 1
- HXWALXSAVBLTPK-NUTKFTJISA-N Leu-Ala-Trp Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CC(C)C)N HXWALXSAVBLTPK-NUTKFTJISA-N 0.000 description 1
- REPPKAMYTOJTFC-DCAQKATOSA-N Leu-Arg-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O REPPKAMYTOJTFC-DCAQKATOSA-N 0.000 description 1
- KKXDHFKZWKLYGB-GUBZILKMSA-N Leu-Asn-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N KKXDHFKZWKLYGB-GUBZILKMSA-N 0.000 description 1
- QLQHWWCSCLZUMA-KKUMJFAQSA-N Leu-Asp-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 QLQHWWCSCLZUMA-KKUMJFAQSA-N 0.000 description 1
- WCTCIIAGNMFYAO-DCAQKATOSA-N Leu-Cys-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O WCTCIIAGNMFYAO-DCAQKATOSA-N 0.000 description 1
- LOLUPZNNADDTAA-AVGNSLFASA-N Leu-Gln-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LOLUPZNNADDTAA-AVGNSLFASA-N 0.000 description 1
- HPBCTWSUJOGJSH-MNXVOIDGSA-N Leu-Glu-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O HPBCTWSUJOGJSH-MNXVOIDGSA-N 0.000 description 1
- XQXGNBFMAXWIGI-MXAVVETBSA-N Leu-His-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CC(C)C)CC1=CN=CN1 XQXGNBFMAXWIGI-MXAVVETBSA-N 0.000 description 1
- AVEGDIAXTDVBJS-XUXIUFHCSA-N Leu-Ile-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AVEGDIAXTDVBJS-XUXIUFHCSA-N 0.000 description 1
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 1
- DSFYPIUSAMSERP-IHRRRGAJSA-N Leu-Leu-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N DSFYPIUSAMSERP-IHRRRGAJSA-N 0.000 description 1
- UCBPDSYUVAAHCD-UWVGGRQHSA-N Leu-Pro-Gly Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UCBPDSYUVAAHCD-UWVGGRQHSA-N 0.000 description 1
- IWMJFLJQHIDZQW-KKUMJFAQSA-N Leu-Ser-Phe Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 IWMJFLJQHIDZQW-KKUMJFAQSA-N 0.000 description 1
- SVBJIZVVYJYGLA-DCAQKATOSA-N Leu-Ser-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O SVBJIZVVYJYGLA-DCAQKATOSA-N 0.000 description 1
- AEDWWMMHUGYIFD-HJGDQZAQSA-N Leu-Thr-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O AEDWWMMHUGYIFD-HJGDQZAQSA-N 0.000 description 1
- KLSUAWUZBMAZCL-RHYQMDGZSA-N Leu-Thr-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(O)=O KLSUAWUZBMAZCL-RHYQMDGZSA-N 0.000 description 1
- WGAZVKFCPHXZLO-SZMVWBNQSA-N Leu-Trp-Glu Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N WGAZVKFCPHXZLO-SZMVWBNQSA-N 0.000 description 1
- ZZHPLPSLBVBWOA-WDSOQIARSA-N Lys-Met-Trp Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCCCN)N ZZHPLPSLBVBWOA-WDSOQIARSA-N 0.000 description 1
- TVOOGUNBIWAURO-KATARQTJSA-N Lys-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CCCCN)N)O TVOOGUNBIWAURO-KATARQTJSA-N 0.000 description 1
- UZWMJZSOXGOVIN-LURJTMIESA-N Met-Gly-Gly Chemical compound CSCC[C@H](N)C(=O)NCC(=O)NCC(O)=O UZWMJZSOXGOVIN-LURJTMIESA-N 0.000 description 1
- JACMWNXOOUYXCD-JYJNAYRXSA-N Met-Val-Phe Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JACMWNXOOUYXCD-JYJNAYRXSA-N 0.000 description 1
- 206010054949 Metaplasia Diseases 0.000 description 1
- XZFYRXDAULDNFX-UHFFFAOYSA-N N-L-cysteinyl-L-phenylalanine Natural products SCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XZFYRXDAULDNFX-UHFFFAOYSA-N 0.000 description 1
- AUEJLPRZGVVDNU-UHFFFAOYSA-N N-L-tyrosyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CC1=CC=C(O)C=C1 AUEJLPRZGVVDNU-UHFFFAOYSA-N 0.000 description 1
- 108010066427 N-valyltryptophan Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- CGOMLCQJEMWMCE-STQMWFEESA-N Phe-Arg-Gly Chemical compound NC(N)=NCCC[C@@H](C(=O)NCC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 CGOMLCQJEMWMCE-STQMWFEESA-N 0.000 description 1
- KZRQONDKKJCAOL-DKIMLUQUSA-N Phe-Leu-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KZRQONDKKJCAOL-DKIMLUQUSA-N 0.000 description 1
- GRVMHFCZUIYNKQ-UFYCRDLUSA-N Phe-Phe-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O GRVMHFCZUIYNKQ-UFYCRDLUSA-N 0.000 description 1
- NJJBATPLUQHRBM-IHRRRGAJSA-N Phe-Pro-Ser Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)N)C(=O)N[C@@H](CO)C(=O)O NJJBATPLUQHRBM-IHRRRGAJSA-N 0.000 description 1
- JXQVYPWVGUOIDV-MXAVVETBSA-N Phe-Ser-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JXQVYPWVGUOIDV-MXAVVETBSA-N 0.000 description 1
- MCIXMYKSPQUMJG-SRVKXCTJSA-N Phe-Ser-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MCIXMYKSPQUMJG-SRVKXCTJSA-N 0.000 description 1
- 238000010806 PrimeScriptTM RT Reagent kit Methods 0.000 description 1
- APKRGYLBSCWJJP-FXQIFTODSA-N Pro-Ala-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(O)=O APKRGYLBSCWJJP-FXQIFTODSA-N 0.000 description 1
- IHCXPSYCHXFXKT-DCAQKATOSA-N Pro-Arg-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O IHCXPSYCHXFXKT-DCAQKATOSA-N 0.000 description 1
- GOMUXSCOIWIJFP-GUBZILKMSA-N Pro-Ser-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GOMUXSCOIWIJFP-GUBZILKMSA-N 0.000 description 1
- FDMCIBSQRKFSTJ-RHYQMDGZSA-N Pro-Thr-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O FDMCIBSQRKFSTJ-RHYQMDGZSA-N 0.000 description 1
- FZXSYIPVAFVYBH-KKUMJFAQSA-N Pro-Tyr-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O FZXSYIPVAFVYBH-KKUMJFAQSA-N 0.000 description 1
- 238000012181 QIAquick gel extraction kit Methods 0.000 description 1
- 108010079005 RDV peptide Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- DSSOYPJWSWFOLK-CIUDSAMLSA-N Ser-Cys-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(O)=O DSSOYPJWSWFOLK-CIUDSAMLSA-N 0.000 description 1
- GZSZPKSBVAOGIE-CIUDSAMLSA-N Ser-Lys-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O GZSZPKSBVAOGIE-CIUDSAMLSA-N 0.000 description 1
- MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 description 1
- VGQVAVQWKJLIRM-FXQIFTODSA-N Ser-Ser-Val Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O VGQVAVQWKJLIRM-FXQIFTODSA-N 0.000 description 1
- GSCVDSBEYVGMJQ-SRVKXCTJSA-N Ser-Tyr-Asp Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CO)N)O GSCVDSBEYVGMJQ-SRVKXCTJSA-N 0.000 description 1
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 1
- KCRQEJSKXAIULJ-FJXKBIBVSA-N Thr-Gly-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O KCRQEJSKXAIULJ-FJXKBIBVSA-N 0.000 description 1
- XOWKUMFHEZLKLT-CIQUZCHMSA-N Thr-Ile-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(O)=O XOWKUMFHEZLKLT-CIQUZCHMSA-N 0.000 description 1
- IJVNLNRVDUTWDD-MEYUZBJRSA-N Thr-Leu-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IJVNLNRVDUTWDD-MEYUZBJRSA-N 0.000 description 1
- SPVHQURZJCUDQC-VOAKCMCISA-N Thr-Lys-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(O)=O SPVHQURZJCUDQC-VOAKCMCISA-N 0.000 description 1
- QFEYTTHKPSOFLV-OSUNSFLBSA-N Thr-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H]([C@@H](C)O)N QFEYTTHKPSOFLV-OSUNSFLBSA-N 0.000 description 1
- VEIKMWOMUYMMMK-FCLVOEFKSA-N Thr-Phe-Phe Chemical compound C([C@H](NC(=O)[C@@H](N)[C@H](O)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 VEIKMWOMUYMMMK-FCLVOEFKSA-N 0.000 description 1
- OFNPHOGOJLNVLL-KCTSRDHCSA-N Trp-Ala-His Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CNC3=CC=CC=C32)N OFNPHOGOJLNVLL-KCTSRDHCSA-N 0.000 description 1
- WMBFONUKQXGLMU-WDSOQIARSA-N Trp-Leu-Val Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N WMBFONUKQXGLMU-WDSOQIARSA-N 0.000 description 1
- UQHPXCFAHVTWFU-BVSLBCMMSA-N Trp-Phe-Val Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O UQHPXCFAHVTWFU-BVSLBCMMSA-N 0.000 description 1
- ABRICLFKFRFDKS-IHPCNDPISA-N Trp-Ser-Tyr Chemical compound C([C@H](NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)C(O)=O)C1=CC=C(O)C=C1 ABRICLFKFRFDKS-IHPCNDPISA-N 0.000 description 1
- WBZOZLNLXVBCNW-LTHWPDAASA-N Trp-Thr-Ile Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(O)=O)[C@@H](C)O)=CNC2=C1 WBZOZLNLXVBCNW-LTHWPDAASA-N 0.000 description 1
- SWSUXOKZKQRADK-FDARSICLSA-N Trp-Val-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N SWSUXOKZKQRADK-FDARSICLSA-N 0.000 description 1
- ARJASMXQBRNAGI-YESZJQIVSA-N Tyr-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N ARJASMXQBRNAGI-YESZJQIVSA-N 0.000 description 1
- VXFXIBCCVLJCJT-JYJNAYRXSA-N Tyr-Pro-Pro Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N1CCC[C@H]1C(O)=O VXFXIBCCVLJCJT-JYJNAYRXSA-N 0.000 description 1
- FZSPNKUFROZBSG-ZKWXMUAHSA-N Val-Ala-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O FZSPNKUFROZBSG-ZKWXMUAHSA-N 0.000 description 1
- WOCYUGQDXPTQPY-FXQIFTODSA-N Val-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](C(C)C)N WOCYUGQDXPTQPY-FXQIFTODSA-N 0.000 description 1
- LIQJSDDOULTANC-QSFUFRPTSA-N Val-Asn-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N LIQJSDDOULTANC-QSFUFRPTSA-N 0.000 description 1
- LJSZPMSUYKKKCP-UBHSHLNASA-N Val-Phe-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=CC=C1 LJSZPMSUYKKKCP-UBHSHLNASA-N 0.000 description 1
- CKTMJBPRVQWPHU-JSGCOSHPSA-N Val-Phe-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)O)N CKTMJBPRVQWPHU-JSGCOSHPSA-N 0.000 description 1
- BCBFMJYTNKDALA-UFYCRDLUSA-N Val-Phe-Phe Chemical compound N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O BCBFMJYTNKDALA-UFYCRDLUSA-N 0.000 description 1
- GQMNEJMFMCJJTD-NHCYSSNCSA-N Val-Pro-Gln Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O GQMNEJMFMCJJTD-NHCYSSNCSA-N 0.000 description 1
- VIKZGAUAKQZDOF-NRPADANISA-N Val-Ser-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O VIKZGAUAKQZDOF-NRPADANISA-N 0.000 description 1
- CEKSLIVSNNGOKH-KZVJFYERSA-N Val-Thr-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C)C(=O)O)NC(=O)[C@H](C(C)C)N)O CEKSLIVSNNGOKH-KZVJFYERSA-N 0.000 description 1
- AEFJNECXZCODJM-UWVGGRQHSA-N Val-Val-Gly Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@@H](C(C)C)C(=O)NCC([O-])=O AEFJNECXZCODJM-UWVGGRQHSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 1
- 108010078114 alanyl-tryptophyl-alanine Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 108010069926 arginyl-glycyl-serine Proteins 0.000 description 1
- 108010038850 arginyl-isoleucyl-tyrosine Proteins 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000004040 coloring Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000002537 cosmetic Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 108010016616 cysteinylglycine Proteins 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 108010054812 diprotin A Proteins 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 108010078274 isoleucylvaline Proteins 0.000 description 1
- 108010076756 leucyl-alanyl-phenylalanine Proteins 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000015689 metaplastic ossification Effects 0.000 description 1
- 108010005942 methionylglycine Proteins 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 210000003463 organelle Anatomy 0.000 description 1
- 150000002894 organic compounds Chemical class 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000001151 other effect Effects 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 108010065135 phenylalanyl-phenylalanyl-phenylalanine Proteins 0.000 description 1
- 108010012581 phenylalanylglutamate Proteins 0.000 description 1
- 150000004291 polyenes Chemical class 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 108010004914 prolylarginine Proteins 0.000 description 1
- 108010053725 prolylvaline Proteins 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 238000004064 recycling Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000007127 saponification reaction Methods 0.000 description 1
- 238000012772 sequence design Methods 0.000 description 1
- 108010048818 seryl-histidine Proteins 0.000 description 1
- 108010071207 serylmethionine Proteins 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 241000894007 species Species 0.000 description 1
- UNFWWIHTNXNPBV-WXKVUWSESA-N spectinomycin Chemical compound O([C@@H]1[C@@H](NC)[C@@H](O)[C@H]([C@@H]([C@H]1O1)O)NC)[C@]2(O)[C@H]1O[C@H](C)CC2=O UNFWWIHTNXNPBV-WXKVUWSESA-N 0.000 description 1
- 229960000268 spectinomycin Drugs 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 150000003505 terpenes Chemical class 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- 108010078580 tyrosylleucine Proteins 0.000 description 1
- 208000019553 vascular disease Diseases 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/37—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi
- C07K14/39—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from fungi from yeasts
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
- C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P23/00—Preparation of compounds containing a cyclohexene ring having an unsaturated side chain containing at least ten carbon atoms bound by conjugated double bonds, e.g. carotenes
Abstract
本发明公开了一种基因RKcrtYB，其是从红冬孢酵母（Rhodosporidium kratochvilovae）YM25235中分离得到，该基因的核苷酸序列如SEQ ID NO:1所示，该基因编码的氨基酸序列如EDQ ID NO:2所示；本发明将该基因连接到载体上转入到YM25235中，其翻译出的蛋白参与β‑类胡萝卜素合成，该基因可以促使红冬孢酵母YM25235提高合成β‑类胡萝卜素，为大规模商业化生产β‑类类胡萝卜素奠定基础。
Description
技术领域
本发明属于生物技术领域中的微生物基因工程领域，具体涉及从红冬孢酵母（Rhodosporidium kratochvilovae）YM25235中克隆出的RKcrtYB基因，再将该基因连接到载体上转入到YM25235中，利用其调节YM25235中类胡萝卜素合成中关键酶的合成，进而可以大量产生β-类胡萝卜素，为高产量优品质β-类胡萝卜素的工业化生产提供参考。
背景技术
类胡萝卜素（carotenoid）是一类广泛存在于自然界的多烯类化合物，由植物和某些光合微生物生成的呈红色、橘黄色或黄色的亲脂性异戊二烯类色素，由于分子结构中类异戊二烯宫二结构的存在，吸光性较强，所以一般在波长430～570nm之间有吸收峰。目前人们在自然界中已经发现了700多种类胡萝卜素。
类胡萝卜素是一种大分子量的有机化合物，典型结构特点是高度不饱和，所以一般不溶于水，但易溶于大部分有机溶剂中，如丙酮、乙酸乙酯、三氯甲烷等。其最显著特征为含有3～15个共轭双键，共轭双键存在的位置也决定类胡萝卜素的功能特性，包括在光合生物中可起到光吸收和光保护的作用；可作为细胞膜的抗氧化保护剂同等。从细胞水平看，类胡萝卜素主要来源于亚细胞的细胞器(质)，细胞通过增加类胡萝卜素的合成有消除胞内氧自由基的作用。所以类胡萝卜素有很多对人体健康有意义的功能，它能有效降低癌症的发病率，以及抗氧化、预防夜盲症、着色等功能。例如，它作为摄入人体的一种抗氧化剂，能起到有效清除人体内多余的自由基的功效，同时保护人体组织细胞避免受到氧和自由基的损伤，并且作为最安全的着色剂之一，类胡萝卜素广泛用于食品、化妆品、水产养殖业；该色素与人类健康的关系近年来更是倍受关注，人体视觉的形成、上皮细胞的正常生长和分化、生殖均与类胡萝卜素有密切关系，而其独特的抗氧化活性，在减少癌症的发生和发展，降低心血管疾病、白内障的发病率等功效也引起了医药领域极大的关注。
类胡萝卜素在含氧、高温、光照、强酸的条件下极不稳定，容易变质降解或者异构化，通常的保存方式是隔绝空气、低温、避光储存，由于它在碱性环境中比较稳定，因此工业上常用碱性皂化处理的方式来提取类胡萝卜素，这样提取的类胡萝卜素虽然成本小，但生物活性低，进而这样提取类胡萝卜素的产量和质量远远不能满足人们的需求；因此从自然界筛选性状优良的微生物进行类胡萝卜素的生物合成以替代化学合成是近年来的一个研究热点。所以我们希望可以从生物工程方面，能通过基因技术、物种发现、杂交培育等方式，得到高含量的原料，这样必然大大缓解天然类胡萝卜素的生产发展局限。
发明内容
本发明的目的是提供一种基因RKcrtYB，它是从红冬孢酵母（Rhodosporidium kratochvilovae）YM25235中分离得到，其翻译出的蛋白参与β-类胡萝卜素合成，该基因可以促使红冬孢酵母YM25235提高合成β-类胡萝卜素；该基因核苷酸序列如SEQ ID NO:1所示，该基因序列长为1785bp（碱基），该基因编码的氨基酸序列如SEQ ID NO:2所示的多肽，共编码594个氨基酸。
本发明另一目的是提供一种含有基因RKcrtYB的重组表达载体，是将SEQ ID NO:1所示基因直接与原核表达载体pMD18-T连接构建重组载体pMD18-T-RKcrtYB，然后将SEQ IDNO:1所示基因直接与穿梭质粒pRH2034连接构建重组载体pRHRKcrtYB。
本发明另一目的是将基因RKcrtYB应用在生产β-类胡萝卜素中。
本发明从红冬孢酵母（Rhodosporidium kratochvilovae）YM25235的总RNA基因中分离得到调控红冬孢酵母产生类胡萝卜素的基因RKcrtYB，该基因全长1785bp；红冬孢酵母YM25235中RKcrtYB基因的过表达会引起细胞内这个基因的转录水平在一定程度上有一定的提高，继而翻译成相应蛋白，引起红冬孢酵母YM25235中β-类胡萝卜素合成量的提高。本研究结果有助于阐明红冬孢酵母YM25235中产β-类胡萝卜素机制，为揭示微生物提高β-类胡萝卜素产量机制提供参考，将有助于通过基因工程手段对其进行改造来提高β-类胡萝卜素含量，对类胡萝卜素的工业化生产提供良好的应用前景和经济效益，为大规模商业化生产β-类胡萝卜素奠定基础。
附图说明
图1为本发明的红冬孢酵母YM25235的RKcrtYB基因PCR扩增图；
图2 重组质粒pRHRKcrtYB限制性酶切分析1.DNA 分子量标记DL10000；2.空质粒pRH2304的NcoⅠ、EcoRⅤ双酶切；3.重组质粒pRHRKcrtYB的NcoⅠ、EcoRⅤ双酶切；4.RKcrtYB基因的PCR产物；5.DNA 分子量标记DL2000；
图3为重组质粒pRHRKcrtYB的质粒图谱；
图4 重组质粒pRHRKcrtYB转化至红冬孢酵母YM25235菌株阳性克隆验证，1：DNA分子量标记DL5000；2：野生型菌株特异性基因条带；3：重组菌株特异性基因条带；4：特异性基因的cDNA条带；5： DNA分子量标记DL2000；
图5为28℃培养条件下RKcrtYB基因过表达对YM25235菌株类胡萝卜素含量变化的影响：A图是含空质粒pRH2304菌株总类胡萝卜素样品液相图谱；B是含有过表达质粒pRHRKcrtYB菌株总类胡萝卜素样品液相图谱；
图6为两株转基因菌株中β-类胡萝卜素含量结果对比图。
具体实施方式
下面结合附图和实施例对本发明作进一步详细说明，但本发明保护范围不局限于所述内容，实施例中使用的试剂和方法，如无特殊说明，均采用常规试剂和使用常规方法。
实施例1：红冬孢酵母（Rhodosporidium kratochvilovae）YM25235的基因RKcrtYB的克隆
采用OMEGA试剂盒E.Z.N.A Fungal RNA Kit提取红冬孢酵母YM25235总RNA，按照TaKaRa公司试剂盒PrimeScript ®RT reagent Kit With gDNA Eraser(Perfect RealTime)说明书进行，反转录反应合成cDNA，取0.5μL为模板进行聚合酶链式反应，根据转录组测序中发现的RKcrtYB序列；根据红冬孢酵母YM25235的转录组序列设计特异性引物（引物1和引物2）进行PCR扩增；反应所用引物、组份和扩增条件如下：
引物1：RKcrtYB-F：5’-CATGCCATGGATGGGCGGACTGGAC-3’（SEQ ID NO:3）
引物2：RKcrtYB-R：5’-CTCGATATCTCACAGCGCCCTCCACA-3’（SEQ ID NO:4）
（CCATGG为NcoⅠ酶切位点，GATATC为EcoRV酶切位点）
PCR扩增体系如下（50 μL）：
PCR扩增条件为：95℃预变性5 min，再用95℃变性30 s，60℃退火30 s，72℃延伸90 s进行30个循环，最后72℃彻底延伸5 min；反应完后取产物1μL，然后在浓度为1%的琼脂糖凝胶中，进行电泳分析，结果如图1所示，经凝胶成像***成像确认1700bp大小的片段，然后用琼脂糖凝胶DNA回收试剂盒（北京索莱宝科技有限公司）回收目的片段，然后将PCR扩增得到的目的基因连接到pMD18-T上，连接产物转化至大肠杆菌DH5α感受态细胞中，用含有氨苄青霉素（Amp+）的LB固体平板进行筛选，挑取平板上的转化子进行菌落PCR筛选阳性克隆，然后送去昆明硕擎生物科技有限公司测序；测序结果表明，获得一段1785bp长的序列，编码594个氨基酸，命名为RKcrtYB，序列组成如SEQ ID NO:1所示的核苷酸序列，编码SEQ ID NO:2所示氨基酸序列。
实施例2：RKcrtYB过表达重组表达质粒pRHRKcrtYB的构建
采用实施例1中cDNA为模板进行PCR扩增，反应所用引物、反应体系和扩增条件如下：
引物1：RKcrtYB-F：5’-CATGCCATGGATGGGCGGACTGGAC-3’（SEQ ID NO:3）
引物2：RKcrtYB-R：5’-CTCGATATCTCACAGCGCCCTCCACA-3’（SEQ ID NO:4）
（CCATGG为NcoⅠ酶切位点，GATATC为EcoRV酶切位点）
PCR扩增体系如下（50 μL）：
质粒pRH2304双酶切体系如下(50μL)：
KcrtYB片段双酶切体系如下(50μL)：
PCR扩增条件为： 95℃预变性5 min，再用95℃变性30 s，60℃退火30 s，72℃延伸90 s进行30个循环，最后72℃彻底延伸5 min，将上述PCR获得的基因片段和表达载体pRH2034分别用NcoⅠ和EcoRV核酸内切酶进行双酶切，两组酶切体系置于37℃酶切8h，用1%的琼脂糖凝胶电泳检测，再用凝胶回收试剂盒对酶切产物进行纯化和回收；再将回收片段以5:4比例用T4DNA连接酶连接4h，连接产物转入大肠杆菌DH5α感受态细胞中；用含有壮观霉素（Spe+）的LB固体平板进行筛选，挑取平板上的转化子进行菌落PCR，筛选阳性克隆，同时进一步对转化子进行双酶切验证分析，结果显示，如图2第3泳道所示，用NcoI和EcoRV双酶切，重组质粒产生两条带，小分子条带与泳道4中该基因的PCR产物大小一致，大分子条带与泳道2中用相同酶切质粒pRH2034后产生的条带大小一致，这表明所构建的重组质粒正确，进一步测序分析也证明这一点。将重组表达质粒命名为pRHRKcrtYB，该质粒图谱如图3所示。
实施例3：RKcrtYB基因过表达对红冬孢酵母YM25235中β-胡萝卜素合成的影响
1、农杆菌介导转化红冬孢酵母YM25235
采用农杆菌介导转化法将重组质粒pRHRKcrtYB转化至红冬孢酵母YM25235菌株，以含潮霉素B（HygromycinB）终浓度为150 µg/mL的YPD培养基筛选转化子；然后按照上海生工生物工程股份有限公司DNA 提取试剂盒说明书中步骤提取酵母转化子的基因组 DNA，然后进行PCR验证如图4所示；从图4可知RKcrtYB序列已经整合到YM25235基因组中，泳道3可以看出PCR扩增可以获得两条带，一条是菌株自身包含内含子序列的基因组片段，一条带是转化整合的cDNA编码序列片段，未转化RKcrtYB基因的对照菌株YM25235中只扩增得到包含内含子序列的基因组片段（泳道2）。
2、RKcrtYB基因过表达与红冬孢酵母中β-胡萝卜素含量关系的分析
将含pRHRKcrtYB的过表达菌株在28℃条件下培养，提取总类胡萝卜素，并测定其中β-胡萝卜素的含量，以转入空质粒pRH2304的红冬孢酵母菌株为对照（图5A），进行高压液相分析，液相图谱如图5所示；两株转基因菌株中β-胡萝卜素含量结果对比如图6所示；结果表明，转基因菌株YM25235/pRHRKcrtYB的β-胡萝卜素含量相较于对照菌株YM25235/pRH2304的β-胡萝卜素含量明显增加，提高2.23倍。因此，在红冬孢酵母YM25235菌株中过表达RKcrtYB基因会引起β-胡萝卜素增加；即说明RKcrtYB核酸序列确实与红冬孢酵母中类胡萝卜素合成有关。
序列表
<110> 昆明理工大学
<120> 一种基因RKcrtYB及其应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1785
<212> DNA
<213> 红冬孢酵母(Rhodosporidium kratochvilovae)
<400> 1
atgggcggac tggactactg gctcgtccat gctcgctgga cgattccccc ggcactcgcc 60
ctctggctcg gctttcgcaa gctgaggacc tggagggacg tctacaagac ttgcttcctc 120
atcaccatcg ctgtaacggc gaccataccc tgggactcgt acctcatccg gaaccgcatc 180
tggtcatacc ccgactcgtc cgtcgtcggc cccaccctct tctccatccc ctacgaggag 240
atcttcttct tctttgtgca aacgtacatc acctcgaccc tctacgccgt cctcactcgc 300
cctatcgtcc atgccgtcct cctgccgcgc acgcccagcg aggggcgctc cgtgcgctgg 360
acggggacgg ccttgttggc ggccgtcttc gcgctcgcat gggcgaagct cgcggagggc 420
ggcgagggga cctacctcgc ccttatcgtc gcatgggtcg cgccgttcct cacgctcctt 480
tggttcgtcg cctcgaccca cctccgcgcc atgccgcgtt cgactcttct cctcgccatc 540
ttcatcccga cgttcttctt gtgggagtgt gacgcgaggg cgttgcagcg cgggacgtgg 600
gtcattgagc aggggacgaa gctcgggctg tcgttccgcg gacttgagat tgaagaagcg 660
gtcttcttcc ttctcaccaa cacgatgatc gtgtttggtc tggtcgcttg cgaccactgc 720
ctcgccgttc acgacattcg ctcgtacgac actggccggt cctccgtctt ccctccactg 780
cgcgacttaa tccccatcct cctcaacaac cccgacgaga cgcagcgcca gcggatcgac 840
gacctccgcg cggcaatcgt catcctctcc gtgcactcga agagcttctc cacggcgagc 900
atggtcttcg acgggcgtct gcggctcgac ttgctcgcac tgtacgcctg gtgccgcgtc 960
tgcgatgacc tagtcgacaa cgcctcctcg gtcgccgccg ccgaggccaa catccgcctc 1020
atctcctcct gcctcgacct cctctacccg ccctctttct cctccgcgac ctcgcacccc 1080
acccgcgtac ccagcaaggc gatcgaggcc gccctcccgg ggctgaacga ggccgagcgc 1140
ggctcgttcc gcctcctctc gctcctcccc atcacgcgcc cgccactcga cgagctcttc 1200
gccggtttcg agacggacct ctcgttcctc gccttcgcag gctcgaaggc gggcgagggc 1260
gaggaggaga cgggcccttc gcggatcccg tccgagctgc cgatcaagac ggacgcggac 1320
ctcctcgcgt acgccaacaa cgtcgcgtcg tcggtcgcgg acctgtgcgt gcagctcgtc 1380
tgggcgcact gtgcgacgag cgtcccgcag gaggagcagc gcgcgatcct cgccgccgcg 1440
cgcgagatgg gccaggcgct ccagctcgtc aacatcgcgc gggacgtccc cgccgacctg 1500
cacatcaaca ggatctacct ccccggccgc gggctcgaca cgcccgtctc ggagctgacg 1560
ccggaccggc gcgagctcct ccgccgcgca cgcgagatgg ccgcgcacag ccgcggcgcg 1620
attgagcgcc ttccgcgcga ggcgaggggc gggatccgcg cggcgtgcga cgtctacttg 1680
tcgatcggcg gggcggtcga ccttgcgctg gatgaggggc gggtggacga gcgcgcgagg 1740
gttgcaaggg ggacgagggc ttggaagatg tggagggcgc tgtga 1785
<210> 2
<211> 594
<212> PRT
<213> 红冬孢酵母(Rhodosporidium kratochvilovae)
<400> 2
Met Gly Gly Leu Asp Tyr Trp Leu Val His Ala Arg Trp Thr Ile Pro
1 5 10 15
Pro Ala Leu Ala Leu Trp Leu Gly Phe Arg Lys Leu Arg Thr Trp Arg
20 25 30
Asp Val Tyr Lys Thr Cys Phe Leu Ile Thr Ile Ala Val Thr Ala Thr
35 40 45
Ile Pro Trp Asp Ser Tyr Leu Ile Arg Asn Arg Ile Trp Ser Tyr Pro
50 55 60
Asp Ser Ser Val Val Gly Pro Thr Leu Phe Ser Ile Pro Tyr Glu Glu
65 70 75 80
Ile Phe Phe Phe Phe Val Gln Thr Tyr Ile Thr Ser Thr Leu Tyr Ala
85 90 95
Val Leu Thr Arg Pro Ile Val His Ala Val Leu Leu Pro Arg Thr Pro
100 105 110
Ser Glu Gly Arg Ser Val Arg Trp Thr Gly Thr Ala Leu Leu Ala Ala
115 120 125
Val Phe Ala Leu Ala Trp Ala Lys Leu Ala Glu Gly Gly Glu Gly Thr
130 135 140
Tyr Leu Ala Leu Ile Val Ala Trp Val Ala Pro Phe Leu Thr Leu Leu
145 150 155 160
Trp Phe Val Ala Ser Thr His Leu Arg Ala Met Pro Arg Ser Thr Leu
165 170 175
Leu Leu Ala Ile Phe Ile Pro Thr Phe Phe Leu Trp Glu Cys Asp Ala
180 185 190
Arg Ala Leu Gln Arg Gly Thr Trp Val Ile Glu Gln Gly Thr Lys Leu
195 200 205
Gly Leu Ser Phe Arg Gly Leu Glu Ile Glu Glu Ala Val Phe Phe Leu
210 215 220
Leu Thr Asn Thr Met Ile Val Phe Gly Leu Val Ala Cys Asp His Cys
225 230 235 240
Leu Ala Val His Asp Ile Arg Ser Tyr Asp Thr Gly Arg Ser Ser Val
245 250 255
Phe Pro Ser Leu Arg Asp Phe Ile Pro Ile Leu Leu Asn Asn Pro Asp
260 265 270
Glu Thr Gln Arg Gln Arg Ile Asp Asp Leu Arg Ala Ala Ile Val Ile
275 280 285
Leu Ser Val His Ser Lys Ser Phe Ser Thr Ala Ser Met Val Phe Asp
290 295 300
Gly Arg Leu Arg Leu Asp Leu Leu Ala Leu Tyr Ala Trp Cys Arg Val
305 310 315 320
Cys Asp Asp Leu Val Asp Asn Ala Ser Ser Val Ala Ala Ala Glu Ala
325 330 335
Asn Ile Arg Leu Ile Ser Ser Cys Leu Asp Leu Leu Tyr Pro Pro Ser
340 345 350
Phe Ser Ser Ala Thr Ser His Pro Thr Arg Val Pro Ser Lys Ala Ile
355 360 365
Glu Ala Ala Leu Pro Gly Leu Asn Glu Ala Glu Arg Gly Ser Phe Arg
370 375 380
Leu Leu Ser Leu Leu Pro Ile Thr Arg Pro Pro Leu Asp Glu Leu Phe
385 390 395 400
Ala Gly Phe Glu Thr Asp Leu Ser Phe Leu Ala Phe Ala Gly Ser Lys
405 410 415
Ala Gly Glu Gly Glu Glu Glu Thr Gly Pro Ser Arg Ile Pro Ser Glu
420 425 430
Leu Pro Ile Lys Thr Asp Ala Asp Leu Leu Ala Tyr Ala Asn Asn Val
435 440 445
Ala Ser Ser Val Ala Asp Leu Cys Val Gln Leu Val Trp Ala His Cys
450 455 460
Ala Thr Ser Val Pro Gln Glu Glu Gln Arg Ala Ile Leu Ala Ala Ala
465 470 475 480
Arg Glu Met Gly Gln Ala Leu Gln Leu Val Asn Ile Ala Arg Asp Val
485 490 495
Pro Ala Asp Leu His Ile Asn Arg Ile Tyr Leu Pro Gly Arg Gly Leu
500 505 510
Asp Thr Pro Val Ser Glu Leu Thr Pro Asp Arg Arg Glu Leu Leu Arg
515 520 525
Arg Ala Arg Glu Met Ala Ala His Ser Arg Gly Ala Ile Glu Arg Leu
530 535 540
Pro Arg Glu Ala Arg Gly Gly Ile Arg Ala Ala Cys Gly Val Tyr Leu
545 550 555 560
Ser Ile Gly Gly Ala Val Asp Leu Ala Leu Asp Glu Gly Arg Val Asp
565 570 575
Glu Arg Ala Arg Val Ala Arg Gly Thr Arg Ala Trp Lys Met Trp Arg
580 585 590
Ala Leu
<210> 3
<211> 25
<212> DNA
<213> 人工序列(Artificial)
<400> 3
catgccatgg atgggcggac tggac 25
<210> 4
<211> 26
<212> DNA
<213> 人工序列(Artificial)
<400> 4
ctcgatatct cacagcgccc tccaca 26
Claims (3)
1.一种基因RKcrtYB，其核苷酸序列如SEQ ID NO:1所示，该基因编码的氨基酸序列如SEQ ID NO:2 所示。
2.一种含有权利要求1所述基因RKcrtYB的重组表达载体。
3.权利要求1所述的基因RKcrtYB在生产β-类胡萝卜素中的应用。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811281423.XA CN109609519A (zh) | 2018-10-31 | 2018-10-31 | 一种基因RKcrtYB及其应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811281423.XA CN109609519A (zh) | 2018-10-31 | 2018-10-31 | 一种基因RKcrtYB及其应用 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109609519A true CN109609519A (zh) | 2019-04-12 |
Family
ID=66001893
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811281423.XA Pending CN109609519A (zh) | 2018-10-31 | 2018-10-31 | 一种基因RKcrtYB及其应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109609519A (zh) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108624600A (zh) * | 2018-05-22 | 2018-10-09 | 昆明理工大学 | 锌指转录因子基因RkMsn4的用途 |
| CN111454854A (zh) * | 2020-05-02 | 2020-07-28 | 昆明理工大学 | 一株产虾青素的红冬孢酵母基因工程菌株 |
Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1184159A (zh) * | 1996-12-02 | 1998-06-10 | 霍夫曼-拉罗奇有限公司 | 改进的类胡萝卜素发酵生产 |
| US20090142322A1 (en) * | 2007-11-30 | 2009-06-04 | E.I. Du Pont De Nemours And Company | Coenzyme Q10 Production in a Recombinant Oleaginous Yeast |
| WO2010123868A1 (en) * | 2009-04-22 | 2010-10-28 | Medicine In Need Corporation | Production of provitamin a carotenoids in mushrooms and uses thereof |
| CN103865818A (zh) * | 2012-12-07 | 2014-06-18 | 上海来益生物药物研究开发中心有限责任公司 | 一种产虾青素基因工程菌的构建方法 |
| CN103865817A (zh) * | 2012-12-07 | 2014-06-18 | 上海来益生物药物研究开发中心有限责任公司 | 一种产β-胡萝卜素基因工程菌的构建方法 |
| CN105087406A (zh) * | 2015-07-22 | 2015-11-25 | 天津大学 | 一种重组酵母菌株及其构建方法和应用 |
| CN105779319A (zh) * | 2016-03-23 | 2016-07-20 | 天津大学 | 一种重组酵母菌株及其构建方法和应用 |
| WO2016154314A1 (en) * | 2015-03-23 | 2016-09-29 | Arch Innotek, Llc | Compositions and methods of biosynthesizing carotenoids and their derivatives |
| CN106148417A (zh) * | 2015-03-26 | 2016-11-23 | 清华大学 | 标准化生物元件的设计和构建方法及其应用 |
| CN106987550A (zh) * | 2017-05-18 | 2017-07-28 | 陕西师范大学 | 一种产β‑胡萝卜素的重组菌及其构建方法和应用 |
| WO2017212030A1 (en) * | 2016-06-09 | 2017-12-14 | Deinove | Methods of producing astaxanthin or precursors thereof |
| CN108359616A (zh) * | 2018-02-14 | 2018-08-03 | 天津大学 | Yel013w基因和yer042w基因的应用 |
| CN108395473A (zh) * | 2018-02-13 | 2018-08-14 | 中国农业科学院生物技术研究所 | 植物类胡萝卜素合成相关蛋白及其编码基因与应用 |
| CN109097385A (zh) * | 2018-07-26 | 2018-12-28 | 西安海斯夫生物科技有限公司 | 一种产β-胡萝卜素类球红细菌工程菌株及其构建方法 |
-
2018
- 2018-10-31 CN CN201811281423.XA patent/CN109609519A/zh active Pending
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1184159A (zh) * | 1996-12-02 | 1998-06-10 | 霍夫曼-拉罗奇有限公司 | 改进的类胡萝卜素发酵生产 |
| US20090142322A1 (en) * | 2007-11-30 | 2009-06-04 | E.I. Du Pont De Nemours And Company | Coenzyme Q10 Production in a Recombinant Oleaginous Yeast |
| WO2010123868A1 (en) * | 2009-04-22 | 2010-10-28 | Medicine In Need Corporation | Production of provitamin a carotenoids in mushrooms and uses thereof |
| CN103865818A (zh) * | 2012-12-07 | 2014-06-18 | 上海来益生物药物研究开发中心有限责任公司 | 一种产虾青素基因工程菌的构建方法 |
| CN103865817A (zh) * | 2012-12-07 | 2014-06-18 | 上海来益生物药物研究开发中心有限责任公司 | 一种产β-胡萝卜素基因工程菌的构建方法 |
| WO2016154314A1 (en) * | 2015-03-23 | 2016-09-29 | Arch Innotek, Llc | Compositions and methods of biosynthesizing carotenoids and their derivatives |
| CN106148417A (zh) * | 2015-03-26 | 2016-11-23 | 清华大学 | 标准化生物元件的设计和构建方法及其应用 |
| CN105087406A (zh) * | 2015-07-22 | 2015-11-25 | 天津大学 | 一种重组酵母菌株及其构建方法和应用 |
| CN105779319A (zh) * | 2016-03-23 | 2016-07-20 | 天津大学 | 一种重组酵母菌株及其构建方法和应用 |
| WO2017212030A1 (en) * | 2016-06-09 | 2017-12-14 | Deinove | Methods of producing astaxanthin or precursors thereof |
| CN106987550A (zh) * | 2017-05-18 | 2017-07-28 | 陕西师范大学 | 一种产β‑胡萝卜素的重组菌及其构建方法和应用 |
| CN108395473A (zh) * | 2018-02-13 | 2018-08-14 | 中国农业科学院生物技术研究所 | 植物类胡萝卜素合成相关蛋白及其编码基因与应用 |
| CN108359616A (zh) * | 2018-02-14 | 2018-08-03 | 天津大学 | Yel013w基因和yer042w基因的应用 |
| CN109097385A (zh) * | 2018-07-26 | 2018-12-28 | 西安海斯夫生物科技有限公司 | 一种产β-胡萝卜素类球红细菌工程菌株及其构建方法 |
Non-Patent Citations (5)
| Title |
|---|
| FIRRINCIELI,A.等: ""Rhodotorula graminis WP1 hypothetical protein partial mRNA"", 《GENBANK DATABASE》 * |
| GUO,J.W.: ""lycopene cyclase/phytoene synthase [Rhodotorula diobovata]"", 《GENBANK DATABASE》 * |
| WENPING XIE等: ""Construction of a Controllable β-carotene Biosynthetic Pathway by Decentralized Assembly Strategy in Saccharomyces cerevisiae"", 《CELLULAR AND METABOLIC ENGINEERING》 * |
| 刘小岭: ""红冬孢酵母类胡萝卜素合成关键酶基因Pdh和Lc的研究"", 《中国优秀硕士学位论文全文数据库基础科学辑》 * |
| 陶俊等: ""类胡萝卜素合成的相关基因及其基因工程"", 《生物工程学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108624600A (zh) * | 2018-05-22 | 2018-10-09 | 昆明理工大学 | 锌指转录因子基因RkMsn4的用途 |
| CN111454854A (zh) * | 2020-05-02 | 2020-07-28 | 昆明理工大学 | 一株产虾青素的红冬孢酵母基因工程菌株 |
| CN111454854B (zh) * | 2020-05-02 | 2022-05-06 | 昆明理工大学 | 一株产虾青素的红冬孢酵母基因工程菌株 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109666683B (zh) | 乙酰辅酶A乙酰转移酶基因RKAcaT2及其应用 | |
| JP3782442B2 (ja) | 遺伝子操作した宿主におけるカロテノイドの生合成 | |
| CN109777815B (zh) | HMG-CoA合成酶基因RKHMGCS及其应用 | |
| CN112410355B (zh) | 一种酰基辅酶a氧化酶2基因rkacox2及其应用 | |
| CN110747206B (zh) | 3-羟基-3-甲基戊二酸单酰辅酶a还原酶基因rkhmgr及其应用 | |
| CN111454854B (zh) | 一株产虾青素的红冬孢酵母基因工程菌株 | |
| CN107058348B (zh) | 一种提高植物赤霉病抗性的小麦基因及其应用 | |
| CN113621630B (zh) | 3-酮脂酰-CoA硫解酶基因RkACAA1-1及其应用 | |
| CN109536518A (zh) | 一种八氢番茄红素脱氢酶基因RKcrtI及其应用 | |
| CN115011616B (zh) | 一种乙醛脱氢酶基因rkaldh及其应用 | |
| CN108503699A (zh) | 枸杞基因以及其编码蛋白质、重组载体、及其用途 | |
| CN112724217B (zh) | 一种青蒿MYB类转录因子AaMYB108及其应用 | |
| CN109609519A (zh) | 一种基因RKcrtYB及其应用 | |
| CN101580543A (zh) | 植物调节花色素苷合成的转录因子-myb编码基因 | |
| CN109748959B (zh) | 花青素合成相关蛋白SlANT1L及其编码基因与应用 | |
| CN105566463A (zh) | 一种与叶绿素合成相关的蛋白及其编码基因与应用 | |
| CN101413006B (zh) | 一种干旱诱导的水稻花特异性启动子及其应用 | |
| CN108624600B (zh) | 锌指转录因子基因RkMsn4的用途 | |
| CN109943575B (zh) | 黄芩花青素转录调控因子SbMYB75和SbDEL的基因克隆、载体构建及其应用 | |
| CN108823178B (zh) | 大黄素糖基转移酶蛋白FtUGT73BE5及其编码基因与应用 | |
| CN107828749B (zh) | 一种深海海参来源的耐低温超氧化物歧化酶MnSOD及其制备方法 | |
| CN112851779B (zh) | 一种培育花青苷含量提高的转基因植物的方法 | |
| CN106701800B (zh) | 一种出芽短梗霉聚酮合成酶基因及其应用 | |
| CN115925848A (zh) | 一种石斛ERF类转录因子基因DoERF5及其应用 | |
| CN104278015A (zh) | 一株高效过表达内源虾青素合成酶基因的法夫酵母菌株 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20190412 |