CN109439778B - Mark primer pair of lactobacillus rhamnosus hsryfm1301 derived from Bama longevity village, identification method and application thereof - Google Patents
Mark primer pair of lactobacillus rhamnosus hsryfm1301 derived from Bama longevity village, identification method and application thereof Download PDFInfo
- Publication number
- CN109439778B CN109439778B CN201811533437.6A CN201811533437A CN109439778B CN 109439778 B CN109439778 B CN 109439778B CN 201811533437 A CN201811533437 A CN 201811533437A CN 109439778 B CN109439778 B CN 109439778B
- Authority
- CN
- China
- Prior art keywords
- hsryfm1301
- lactobacillus rhamnosus
- hsryfm
- dna
- identification method
- 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.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to the technical field of biology, in particular to a marker primer pair of lactobacillus rhamnosus hsryfm1301 originated from Bama Changshoucun. The invention also discloses a method for identifying the lactobacillus rhamnosus hsryfm 1301. The invention also discloses application of the identification method in the verification of the existence of lactobacillus rhamnosus hsryfm1301 in a fermented product. The invention also discloses application of the identification method in the verification of the existence of lactobacillus rhamnosus hsryfm1301 in excrement. The invention also discloses application of the identification method in strain counting of the lactobacillus rhamnosus homogeneous and heterogeneous mixed culture. The primer pair designed by the invention can be amplified to a stable band from lactobacillus rhamnosus hsryfm1301, forms a specific map, has stable amplification capacity, and can be used as a marker primer of lactobacillus rhamnosus hsryfm1301 strain.
Description
Technical Field
The invention relates to the technical field of biology, in particular to PCR strain identification and application of probiotic lactobacillus rhamnosus hsryfm1301 originated from Guangxi Bama longevity village.
Technical Field
With the development of economic society and the acceleration of life rhythm, more and more people are in a sub-health state or suffer from 'rich diseases'. In recent years, a large number of researches show that lactic acid bacteria are important probiotics in human intestinal tracts, play important roles in maintaining the micro-ecological balance of host intestinal tracts, improving the metabolic function and the immune system function of human bodies, and can relieve the sub-health state of the human bodies and even treat partial diseases. However, the fermentation performance, intestinal tolerance and probiotic function of different strains of the same strain of lactic acid bacteria are greatly different, and the function research of the probiotics needs to reach the level of the strains. The probiotic lactobacillus rhamnosus hsryfm1301 originated from Guangxi Bama longevity village has prominent probiotic effect, but because the identification of the strain is complicated and difficult, a quick and effective means for quickly confirming the strain is still lacked. The invention designs 4 pairs of marker primers capable of forming a specific amplification map on lactobacillus rhamnosus hsryfm1301 by utilizing multi-gene group comparison, can quickly confirm and identify lactobacillus rhamnosus hsryfm1301, and provides a thought for quickly identifying other strains.
Disclosure of Invention
The invention aims to provide a marker primer capable of rapidly identifying lactobacillus rhamnosus hsryfm1301 and application thereof, and the primer pair designed by the invention can be amplified to a stable strip from lactobacillus rhamnosus hsryfm1301, forms a specific map, has stable amplification capacity, and can be used as a marker primer of lactobacillus rhamnosus hsryfm1301 strain.
The invention designs 4 pairs of marking primers of lactobacillus rhamnosus hsryfm1301 through multi-strain whole genome gene family analysis (the primer characteristics are shown in table 1). PCR amplification proves that the 4 pairs of primers can form a specific amplification map for the lactobacillus rhamnosus hsryfm1301, have the capability of confirming the lactobacillus rhamnosus hsryfm1301 in a large number of strains, and can be used as a marker primer for the lactobacillus rhamnosus hsryfm 1301.
TABLE 1 identifying primer Properties
The invention also discloses an identification method of lactobacillus rhamnosus hsryfm1301, which comprises the following steps:
(1) extracting DNA in a sample to be detected;
(2) amplifying DNA of a sample to be detected by using rTaq DNA polymerase by using a marker primer pair of lactobacillus rhamnosus hsryfm1301, and carrying out electrophoresis on a PCR product by using 1% agarose gel;
(3) observation after EB staining:
if hsryfm1301-1F/R can amplify a band size of 900bp from the DNA of the sample to be tested, hsryfm1301-2F/R can amplify a band size of 1100bp from the DNA of the sample to be tested, hsryfm 1301-3F/R can amplify a band size of 1000bp from the DNA of the sample to be tested, and hsryfm 1301-4F/R can amplify a band size of 800bp from the DNA of the sample to be tested, the sample is proved to contain Lactobacillus rhamnosus hsryfm 1301.
Preferably, the amplification conditions are set forth in Table 2:
TABLE 24 amplification conditions and procedures for the primers
Preferably, the electrophoresis conditions are: 120V,300mA,30 min.
The invention also discloses application of the identification method in the verification of the existence of lactobacillus rhamnosus hsryfm1301 in a fermented product.
The invention also discloses application of the identification method in the verification of the existence of lactobacillus rhamnosus hsryfm1301 in excrement.
The invention also discloses application of the identification method in strain counting of the lactobacillus rhamnosus homogeneous and heterogeneous mixed culture.
Compared with the prior art, the invention has the following beneficial effects:
firstly, carrying out whole genome sequencing on lactobacillus rhamnosus hsryfm1301 and LV108 which are separated from Guangxi Bama longevity village and have intestinal environment tolerance capacity and probiotic functions, and carrying out gene family analysis on the 2 genome and lactobacillus rhamnosus GG and Lc705 genomes to obtain a specific gene of lactobacillus rhamnosus hsryfm 1301; 4 pairs of primers are designed according to the sequence of the unique gene, and the capability of rapidly identifying the strain of lactobacillus rhamnosus hsryfm1301 is evaluated. The results show that the use of 4 pairs of primers hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R can amplify a distinct specific band from Lactobacillus rhamnosus hsryfm 1301.
Secondly, amplifying DNA of lactobacillus plantarum, lactobacillus helveticus, lactobacillus delbrueckii subsp bulgaricus, lactobacillus casei, lactobacillus fermentum, streptococcus thermophilus and other 16 strains of lactobacillus rhamnosus by using 4 pairs of primers, wherein the obtained amplification maps are different from hsryfm 1301; the 4 pairs of primers were all able to amplify a band from lactobacillus rhamnosus hsryfm1301 in production. The results show that hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R can be amplified to a stable band from lactobacillus rhamnosus hsryfm1301, form a specific map, have stable amplification capacity and can be used as a marker primer of lactobacillus rhamnosus hsryfm1301 strain. The primer can be used for identifying the lactobacillus rhamnosus hsryfm1301 from the mixture, does not need complex instruments, and can greatly save the operation time and the cost.
Drawings
FIG. 1 is a PCR amplification map of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R against the mother strain of Lactobacillus rhamnosus hsryfm1301 and the reference strain LGG.
FIG. 2a-1 is a PCR amplification profile of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R on a non-homologous lactic acid bacteria;
FIGS. 2a-2 are PCR amplification profiles of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R for non-homologous lactic acid bacteria;
FIG. 2b is a PCR amplification profile of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R against different strains of Lactobacillus rhamnosus;
in the figure: LV108 was the reference strain for primer design, belonging to Lactobacillus rhamnosus; LV-1, R13, R26 and LPRA-1 are all different lactobacillus rhamnosus strains;
FIG. 2c is a PCR amplification profile of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R against different strains of Lactobacillus rhamnosus.
In the figure: sp1, 1505, LY0, BG, Bm03 are all different strains of Lactobacillus rhamnosus;
FIG. 2d is a PCR amplification profile of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R against different strains of Lactobacillus rhamnosus.
In the figure: grx19, Bm01, F and grx10 are all different strains of lactobacillus rhamnosus;
FIG. 2e is a PCR amplification map of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R against Lactobacillus rhamnosus HN 001;
in the figure: HN001 is Lactobacillus rhamnosus isolated from commercial bacterial powder product (Prodeo);
FIG. 3a is a PCR amplification map of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R against a Lactobacillus rhamnosus hsryfm1301 production strain and product;
FIG. 3b is a PCR amplification map of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R against a Lactobacillus rhamnosus hsryfm1301 production strain and product;
in fig. 3a, 3 b: sample a is hsryfm1301 bacterial powder product (Saccharaceae); b is a mixed strain fermented milk enrichment culture (royal dairy industry under the first day), and c, d and e are strains obtained by purifying fermented milk by using a selective medium.
Detailed Description
The invention screens out the peculiar gene of lactobacillus rhamnosus hsryfm1301 by multi-strain gene family analysis, designs primers (hsryfm 1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R) in the peculiar gene, and verifies the effectiveness of the 4 pairs of primers by PCR amplification. The formation of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R on Lactobacillus rhamnosus hsryfm1301 is clearly distinguished from other strains by PCR amplification experiments on non-homologous lactic acid bacteria (Lactobacillus plantarum, Lactobacillus delbrueckii subspecies bulgaricus, Lactobacillus helveticus, Lactobacillus casei, Lactobacillus fermentum, Streptococcus thermophilus) and homologous different strains (LV108, LV-1, R13, R26, LPRA-1, grx19, sp1, 1505, LY0, BG, Bm03, Bm01, F and grx 10).
1 primer Synthesis and stock test
Carrying out whole genome sequencing on the lactobacillus rhamnosus hsryfm1301, screening specific genes of the lactobacillus rhamnosus hsryfm1301 by taking LGG, Lc705 and LV08 as reference strains through multi-strain gene family analysis, and designing primers in the genes: hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R. The 4 pairs of primers were synthesized by Competition Bioengineering, Inc. (Shanghai). Streaking and purifying a lactobacillus rhamnosus hsryfm1301 mother strain in an MRS solid culture medium, picking a single colony to the MRS culture medium for 24 hours at 37 ℃, transferring to a fresh MRS culture medium (2% v/v), and culturing for 24 hours at 37 ℃; the cells were collected, and DNA of the mother strain of Lactobacillus rhamnosus hsryfm1301 was extracted using a column-type DNA extraction kit (Biotechnology, Shanghai). The DNA of the mother strain of Lactobacillus rhamnosus hsryfm1301 was amplified using rTaq DNA polymerase (Takara, Dalian) according to the procedure of Table 3, and the PCR products were electrophoresed on a 1% agarose gel (120V,300mA,30min), and observed after EB staining.
TABLE 34 amplification conditions and procedures for the primers
Each of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R was able to amplify distinct bands from the DNA of the mother strain of Lactobacillus rhamnosus hsryfm1301, the amplified bands having sizes of approximately 900bp, 1100bp, 1000bp and 800bp, respectively, consistent with the predicted lengths in Table 1 (FIG. 1). None of the 4 primers amplified a band from the DNA of the reference strain LGG, indicating that the results of the gene family analysis are reliable and that the 4 primers are available.
2 amplification detection of non-homologous lactic acid bacteria
The DNA extraction and PCR methods were as above. In order to verify the specificity of the 4 pairs of primers, the DNA of other strains of lactic acid bacteria was amplified first, and the results showed that none of the 4 pairs of primers could amplify to bands from the DNA of Lactobacillus plantarum, Lactobacillus fermentum, Lactobacillus helveticus, Lactobacillus bulgaricus and Streptococcus thermophilus (FIGS. 2a-1, 2 a-2); hsryfm1301-1F/R can be amplified from Lactobacillus casei DNA to a band greater than 5000bp, and has no effect on the discrimination effect (FIG. 2a-1, FIG. 2 a-2). The result shows that the amplification maps of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R on several lactic acid bacteria are different from that of lactobacillus rhamnosus hsryfm1301, and the amplification maps have better distinguishing effect on the level of the species.
Amplification detection of different strains of lactobacillus rhamnosus
The invention aims to perform identification among strains, and 4 pairs of primers are used for amplification detection of other 14 strains of lactobacillus rhamnosus which are separated and stored in a laboratory. Lactobacillus rhamnosus LV108 is also a reference strain for primer design, and no amplification band can be obtained by 4 pairs of primers (FIG. 2b), thereby further confirming the effectiveness of gene family analysis. In other strains, 4 pairs of primers cannot be amplified to bands from the DNA of 13 lactobacillus rhamnosus isolated and obtained in a laboratory (FIGS. 2b,2c and 2d), and show better distinguishing effect. To further verify the reliability of the 4 primers, lactobacillus rhamnosus HN001 was isolated from a commercial bacterial powder product (pedile), and no amplified band was found in any of the 4 primers (fig. 2 e). The result shows that the amplification maps of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R on 15 strains of lactobacillus rhamnosus are obviously different from that of lactobacillus rhamnosus hsryfm1301, and the primers can be used as the marker primers of lactobacillus rhamnosus hsryfm 1301.
Amplification detection of 4 rhamnose lactobacillus production strain and product
In order to verify the stability of hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R on the amplification map of lactobacillus rhamnosus hsryfm1301, the DNA of lactobacillus rhamnosus hsryfm1301 in different products is extracted for amplification detection. The results show that hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R can obtain the same map as the mother strain of lactobacillus rhamnosus hsryfm1301 from the DNA amplification of lactobacillus rhamnosus hsryfm1301 powder, fermented milk purification strains and mixed strains (FIG. 3a, FIG. 3b), show that the amplification effect of 4 pairs of primers on hsryfm1301 is not affected by production culture and other strains, and prove that hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R have specificity and stability on the amplification of lactobacillus rhamnosus hsryfm1301 and can be used as a marker primer for verifying lactobacillus rhamnosus hsryfm 1301.
Application examples
1 the primer of the invention is used for rapidly identifying lactobacillus rhamnosus hsryfm 1301.
Culturing and purifying different lactobacillus rhamnosus, extracting DNA, amplifying the DNA by using hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R (see a program in a table 3), and comparing an amplification map with an amplification map of a lactobacillus rhamnosus hsryfm1301 mother strain, wherein the same is lactobacillus rhamnosus hsryfm 1301.
2 the primers of the invention are used to verify the existence of lactobacillus rhamnosus hsryfm1301 in the fermented product.
Collecting microorganisms (which may not be concentrated when the concentration is higher) in the concentrated fermentation product, and performing lysis on the microorganisms; and (3) performing PCR amplification by using the lysate as a template and using hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R (see a program in table 3), so as to obtain an amplification map which is the same as that of a lactobacillus rhamnosus hsryfm1301 mother strain, and the existence of lactobacillus rhamnosus hsryfm1301 in a fermentation product is shown.
3 the presence of lactobacillus rhamnosus hsryfm1301 in the feces was verified using the primers of the invention.
Collecting microorganisms (suspension can be directly cracked when the concentration is higher) in the concentrated excrement, and cracking the microorganisms; and (3) performing PCR amplification by using the lysate as a template and using hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R (see a program in table 3), so as to obtain an amplification map which is the same as that of a lactobacillus rhamnosus hsryfm1301 mother strain, and the existence of lactobacillus rhamnosus hsryfm1301 in the excrement sample is shown.
Enumeration of hsryfm1301 in 4 Lactobacillus rhamnosus Multi-Strain Mixed cultures
Based on hsryfm1301-1F/R, hsryfm1301-2F/R, hsryfm 1301-3F/R and hsryfm 1301-4F/R, a pUC19 recombinant plasmid containing the corresponding gene fragment was used to generate a standard curve. Collecting the microorganisms (which may not be concentrated when the concentration is high) in the mixed culture, and performing lysis on the microorganisms; counting the number of the lactobacillus rhamnosus hsryfm1301 in the mixed sample by using an RT-qPCR technology.
<110> Huang's group of Yangzhou university GmbH
<120> marker primer pair of probiotic lactobacillus rhamnosus hsryfm1301 originated from Bama longevity village, identification method and application thereof
<160>8
SEQ ID NO.1
<210>1
<211>22
<212>DNA
<213> Artificial sequence
<400>1
CTATTGACGA GCAAGCAGGA TC 22
SEQ ID NO.2
<210>2
<211>22
<212>DNA
<213> Artificial sequence
<400>2
TCAGCTGGTA GACGCGTGTG AA 22
SEQ ID NO.3
<210>3
<211>25
<212>DNA
<213> Artificial sequence
<400>3
TTTTAAGAAC TATATTCAGG AAGGC 25
SEQ ID NO.4
<210>4
<211>25
<212>DNA
<213> Artificial sequence
<400>4
AAATACTAGC GACTTAAAAC AGGAG 25
SEQ ID NO.5
<210>5
<211>24
<212>DNA
<213> Artificial sequence
<400>5
ATGACCCACT CTCAGACTAA CACC 24
SEQ ID NO.6
<210>6
<211>25
<212>DNA
<213> Artificial sequence
<400>6
TAGGTCATTT TGAGGACTCC TTTCG 25
SEQ ID NO.7
<210>7
<211>25
<212>DNA
<213> Artificial sequence
<400>7
AACCGTTTAC ACTTACTTTG CCATG 25
SEQ ID NO.8
<210>8
<211>21
<212>DNA
<213> Artificial sequence
<400>8
CGCTGAAAAC CCGCAATTTA C 21
Claims (6)
1. The marker primer pair of lactobacillus rhamnosus hsryfm1301 originated from Bama longevity village is characterized in that the primer pair comprises:
hsryfm 1301-1F：CTATTGACGAGCAAGCAGGATC，
hsryfm 1301-1R：TCAGCTGGTAGACGCGTGTGAA；
hsryfm 1301-2F：TTTTAAGAACTATATTCAGGAAGGC，
hsryfm 1301-2R：AAATACTAGCGACTTAAAACAGGAG；
hsryfm 1301-3F：ATGACCCACTCTCAGACTAACACC，
hsryfm 1301-3R：TAGGTCATTTTGAGGACTCCTTTCG；
hsryfm 1301-4F：AACCGTTTACACTTACTTTGCCATG，
hsryfm 1301-4R：CGCTGAAAACCCGCAATTTAC。
2. the identification method of lactobacillus rhamnosus hsryfm1301 is characterized by comprising the following steps:
(1) extracting DNA in a sample to be detected;
(2) amplifying the DNA of a sample to be detected by using rTaq DNA polymerase by using the primer pair as claimed in claim 1, and performing electrophoresis on a PCR product by using 1% agarose gel;
(3) observation after EB staining:
if hsryfm1301-1F/R can amplify a band size of 900bp from the DNA of the sample to be tested, hsryfm1301-2F/R can amplify a band size of 1100bp from the DNA of the sample to be tested, hsryfm 1301-3F/R can amplify a band size of 1000bp from the DNA of the sample to be tested, and hsryfm 1301-4F/R can amplify a band size of 800bp from the DNA of the sample to be tested, the sample is proved to contain Lactobacillus rhamnosus hsryfm 1301.
3. The method of claim 2, wherein the amplification conditions are set forth in Table 1:
TABLE 14 amplification conditions and procedures for primers
4. The method of claim 2, wherein the electrophoresis conditions are: 120V,300mA,30 min.
5. Use of the identification method according to claim 2 for the verification of the presence of lactobacillus rhamnosus hsryfm1301 in a fermented product.
6. Use of the identification method according to claim 2 for verifying the presence of lactobacillus rhamnosus hsryfm1301 in faeces.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811533437.6A CN109439778B (en) | 2018-12-14 | 2018-12-14 | Mark primer pair of lactobacillus rhamnosus hsryfm1301 derived from Bama longevity village, identification method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811533437.6A CN109439778B (en) | 2018-12-14 | 2018-12-14 | Mark primer pair of lactobacillus rhamnosus hsryfm1301 derived from Bama longevity village, identification method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109439778A CN109439778A (en) | 2019-03-08 |
| CN109439778B true CN109439778B (en) | 2021-06-04 |
Family
ID=65559374
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811533437.6A Active CN109439778B (en) | 2018-12-14 | 2018-12-14 | Mark primer pair of lactobacillus rhamnosus hsryfm1301 derived from Bama longevity village, identification method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109439778B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109811031A (en) * | 2019-03-05 | 2019-05-28 | 扬州大学 | A kind of novel Lactobacillus rhamnosus identification method |
| CN109943655A (en) * | 2019-04-15 | 2019-06-28 | 内蒙古农业大学 | Qualitative and/or quantitative detection Lactobacillus rhamnosus specific primer, kit and method |
| CN110734880B (en) * | 2019-11-19 | 2021-05-18 | 扬州大学 | Lactobacillus plantarum Bama06 derived from Guangxi Bama and having high vitamin B yield and application thereof |
| CN114426939B (en) * | 2022-02-23 | 2023-10-27 | 扬州大学 | Lactobacillus rhamnosus with ochratoxin A removal effect |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101407835A (en) * | 2007-10-12 | 2009-04-15 | 统一企业(中国)投资有限公司 | Genetic marker and method for detecting rhamnose bacterium lacticum |
| CN102146458A (en) * | 2011-01-26 | 2011-08-10 | 中国农业科学院农业资源与农业区划研究所 | Specific primer pair for PCR (Polymerase Chain Reaction) detection and identification of lactobacillus rhamnosus |
| CN103710290A (en) * | 2013-12-31 | 2014-04-09 | 广西皇氏甲天下乳业股份有限公司 | Probiotics lactobacillus rhamnosus hsryfm1301 originated in Guangxi Bama longevity village and application of probiotics lactobacillus rhamnosus hsryfm1301 |
| CN104726553A (en) * | 2014-12-31 | 2015-06-24 | 苏州百源基因技术有限公司 | Specific primers and probes applied to real-time fluorescent quantitative PCR detection of lactobacillus rhamnosus and detection kit |
-
2018
- 2018-12-14 CN CN201811533437.6A patent/CN109439778B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101407835A (en) * | 2007-10-12 | 2009-04-15 | 统一企业(中国)投资有限公司 | Genetic marker and method for detecting rhamnose bacterium lacticum |
| CN102146458A (en) * | 2011-01-26 | 2011-08-10 | 中国农业科学院农业资源与农业区划研究所 | Specific primer pair for PCR (Polymerase Chain Reaction) detection and identification of lactobacillus rhamnosus |
| CN103710290A (en) * | 2013-12-31 | 2014-04-09 | 广西皇氏甲天下乳业股份有限公司 | Probiotics lactobacillus rhamnosus hsryfm1301 originated in Guangxi Bama longevity village and application of probiotics lactobacillus rhamnosus hsryfm1301 |
| CN104726553A (en) * | 2014-12-31 | 2015-06-24 | 苏州百源基因技术有限公司 | Specific primers and probes applied to real-time fluorescent quantitative PCR detection of lactobacillus rhamnosus and detection kit |
Non-Patent Citations (6)
| Title |
|---|
| Genome Instability in Lactobacillus rhamnosus GG;Wilbert Sybesma等;《Applied and Environmental Microbiology》;20130430;第79卷(第7期);第2233-2239页 * |
| Simultaneous discrimination of species and strains in Lactobacillus rhamnosus using species-specific PCR combined with multiplex minisequencing technology;Chien-Hsun Huang等;《Molecular and Cellular Probes》;20150630;第29卷;第531-533页 * |
| 不同来源鼠李糖乳杆菌的随机扩增多态DNA分析;顾瑞霞等;《微生物学报》;20080404;第48卷(第4期);第426-431页 * |
| 应用特异 PCR 快速鉴定微生物肥料中4种乳酸菌;杨小红等;《微生物学通报》;20140420;第41卷(第4期);第674-680页 * |
| 特异性检测植物乳杆菌的多重PCR方法;陈臣等;《南方农业学报》;20130715(第07期);第1076-1080页 * |
| 长寿老人体内分离的鼠李糖乳杆菌RAPD体系优化研究;顾瑞霞等;《食品科学》;20080315(第03期);第268-271页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109439778A (en) | 2019-03-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109439778B (en) | Mark primer pair of lactobacillus rhamnosus hsryfm1301 derived from Bama longevity village, identification method and application thereof | |
| US10041135B2 (en) | Phage insensitive Streptococcus thermophilus | |
| CN111286550A (en) | Specific primer for amplifying lactobacillus paracasei and application thereof | |
| Alimolaei et al. | An efficient DNA extraction method for Lactobacillus casei, a difficult-to-lyse bacterium | |
| CN113564272B (en) | Rapid identification and detection method for cheese lactobacillus in fermented milk | |
| CN107937581B (en) | Amplification primer pair for lactobacillus sequencing, lactobacillus species identification method and application | |
| CN112143820B (en) | Molecular marker, detection primer and detection method for identifying lactobacillus plantarum and lactobacillus pentosus | |
| CN104450941B (en) | Method for detecting lactobacillus casei strain and kit and primer pair thereof | |
| CN116814822A (en) | Identification method and application of bifidobacterium longum subspecies infancy CICC 6069 strain | |
| CN108866220B (en) | Method and detection kit for detecting nasal flora | |
| CN116121415A (en) | Multiplex fluorescence quantitative PCR kit for simultaneously detecting three bifidobacteria, application and detection method | |
| Li et al. | Species attribution and distinguishing strains of Oenococcus oeni isolated from Chinese wines | |
| CN110317891B (en) | Primer group, reagent, kit, application and detection method for detecting lactobacillus rhamnosus LV108 | |
| CN112029884B (en) | Molecular marker, detection primer and detection method for identifying lactobacillus casei group | |
| CN102978293B (en) | Detection method for specifically detecting lactobacillus plantarum and kit therefor | |
| CN104531873B (en) | Method for detecting lactobacillus casei strain and kit and primer pair used by method | |
| CN104531692B (en) | A kind of method and its kit and primer pair for detecting lactobacillus casei bacterial strain | |
| CN103525914B (en) | Method, primer and kit for counting number of live bacteria of lactobacillus plantarum | |
| JP6032737B2 (en) | Method for detecting bifidobacteria | |
| CN106480168B (en) | Method for identifying interspecies relationship and intraspecies relationship of lactobacillus casei | |
| CN104450949B (en) | A kind of specificity molecular marker DNA sequence of Lactobacillus plantarum and application thereof | |
| CN110358851B (en) | Nucleic acid sequence, primer, method and kit for detecting bacillus cereus | |
| CN110592248A (en) | Method for efficiently identifying/screening clostridium butyricum and application thereof | |
| JP2004329161A (en) | Primer set and detection of bacteria | |
| KR20150014042A (en) | Marker composition for identification of Weissella viridescens |
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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |