CN107586854A - A kind of molecular labeling for influenceing duroc waistline character and application - Google Patents
A kind of molecular labeling for influenceing duroc waistline character and application Download PDFInfo
- Publication number
- CN107586854A CN107586854A CN201710852738.4A CN201710852738A CN107586854A CN 107586854 A CN107586854 A CN 107586854A CN 201710852738 A CN201710852738 A CN 201710852738A CN 107586854 A CN107586854 A CN 107586854A
- Authority
- CN
- China
- Prior art keywords
- duroc
- waistline
- character
- pig
- boar
- 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.)
- Granted
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to molecular marker assisted selection and Animal Genetics technical field, and in particular to a kind of molecular labeling for influenceing duroc waistline character and application.The SNP site of the molecular labeling of described influence duroc waistline character corresponds to the 3400805th nucleotide site T on No. 4 chromosomes of international 10.2 version reference sequences of pig genome>C is mutated.It is used to identify that above-mentioned influence Pigs Kidney encloses the primer pair of the molecular labeling of character present invention also offers a kind of, pass through above-mentioned molecular labeling and primer pair, the molecular mark technology of efficiently and accurately can be established, it is applied in the improvement of Duroc boar waistline character inheritance, so as to improve the waistline of offspring pig, and then meat yield is improved, enterprise profit is improved, increases core competitiveness.
Description
Technical field
The invention belongs to molecular marker assisted selection and Animal Genetics technical field, and in particular to one kind influences Du Luo
Gram Pigs Kidney encloses molecular labeling and the application of character.
Background technology
Under market environment based on mainly being evaluated in Chinese pork pig with live hog, the selection of build phenotype is particularly important.
External boar market seldom can judge its value according to build, nevertheless, still can be to the forelimb, hind leg, motion of pig
The appearance traits such as state, posture, the back of the body, linea ventralis carry out comprehensive assessment selection.In most of China live pig in the market, linea ventralis (abdominal circumference,
Waistline) value of pork pig is determined, it directly affects the profit on sales of end product.Because abdominal circumference is big, waistline is small, linea ventralis is uneven
Directly, pregnant, potbelly is commonly called as, this boar causes pork pig finishing loss larger, have impact on boar and finally sell because fat meat is more
The profit gone out.Further, since kind pork tripe sirloin it is larger so as to cause commodity pig farm to be unwilling to introduce a fine variety cause such boar utilization rate drop
Low situation is of common occurrence, and for pig, enterprise brings huge economic loss.Because the selection of build need to take into account it is numerous, very
Hardly possible takes into account waistline while other characters are selected, and difficulty is brought for waistline improvement.The Phenotypic Selection and measure of waistline are not
There are system and in detail research.By the market survey of the country, requirement of the different zones to waistline is different, and this causes to select in boar
When educating, the breeding objective of the appearance traits such as each terminal male strain waistline slightly has difference.Therefore, carry out for market demands to waist
Enclose phenotype and carry out more detailed measure and selection, furthermore it is also possible to be reflected by the analysis of this phenotype test and genome
Not Kong Zhi boar build gene and major gene loci, it is difficult fundamentally to solve waistline improvement using the means of molecular breeding
Topic.
Duroc is one of bacon hogs kind famous in the world.In the breeding of method of modern goods pork pig, Du Luo
Gram used generally as terminal male, coverage is larger.Therefore, the waistline character of duroc is improved, Ke Yiyou
The build of effect ground improvement market pig, improves its selling price.
The content of the invention
In order to overcome deficiency and shortcoming, primary and foremost purpose of the invention existing for boar build seed selection to be in the prior art to carry
For a kind of molecular labeling for influenceing duroc waistline character.
Another object of the present invention is to the application for the molecular labeling for providing above-mentioned influence duroc waistline character.
It is still another object of the present invention to provide a kind of molecular labeling for identifying above-mentioned influence duroc waistline character
Primer pair.
Fourth object of the present invention is the application for providing above-mentioned primer pair.
The 5th purpose of the present invention is a kind of genetic improvement method for providing duroc.
A kind of molecular labeling for influenceing duroc waistline character, its SNP site correspond to international pig reference gene group
T on 10.2 edition 4 chromosomes at 3400805bp>C is mutated；
The SNP site of the molecular labeling of described influence duroc waistline character is (being located at should shown in SEQ ID NO.1
The nucleic acid single base of sequence fragment the 306th is mutated, and is named as：g.306T>C), the M wherein in sequence is T or C, causes waistline character
Difference；
The molecular labeling of described influence duroc waistline character is in identification duroc waistline character and genetic breeding
Middle application；
A kind of method of Duroc boar strain using the high lean meat percentage of above-mentioned molecular selection, is comprised the following steps：
Detect the molecular labeling of above-mentioned influence duroc waistline character on No. 4 chromosomes of pig；Described molecular labeling
SNP site is T or C, eliminates T and retains C；
The boar is preferably purebred American Duroc pig and its synthesis system (Guangdong Wen Shi boars Science and Technology Ltd.)；
A kind of primer pair for being used to identify the molecular labeling of above-mentioned influence duroc waistline character, comprising primer C-F and
C-R, its nucleotide sequence are as follows：
Primer C-F：5’-GGGTGAGGGTGGGATTAG-3’；
Primer C-R：5’-TTGCGTTAGTGATTGTCCTG-3’；
Described primer pair influences to apply in duroc waistline character in identification；
Described primer pair is applied in duroc molecular marker assisted selection breeding；
Described primer pair is applied in the Duroc boar strain for cultivating high lean meat；
The method of the genetic improvement of one boar, is comprised the following steps：
The molecular labeling of the above-mentioned influence duroc waistline character of boar in nucleus herds of breeding pigs is determined, and according to the upper of pig
The genotype of rheme point makes corresponding selection during boar is chosen seeds：The Systematic Breeding world pig reference gene group of boar
CC types individual on 10.2 edition 4 chromosomes at 3400805bp, eliminate the TT types and CT types individual of the point；To be improved by generation
The frequency of the allele C in the site, so as to the waistline of improved progeny pig；
Described boar includes Duroc and its synthesis system；
The present invention is had the following advantages relative to prior art and effect：
(1) present invention studies and determines to influence the molecular labeling of duroc waistline character, verifies it to waistline character
Influential effect, it is final to establish the molecular marker assisted selection breeding technique that quick improvement is carried out to waistline character, significantly improve
Duroc and its build seed selection process of synthesis system, the demand in live hog market is adapted to, the price of pork pig is improved, increases for enterprise
Add profit on sales, improve core competitiveness.
(2) present invention provides a kind of primer pair for being used to identify the molecular labeling for influenceing duroc waistline character, passes through
The molecular labeling and primer pair, the molecular mark technology of efficiently and accurately can be established, fast and accurately build is carried out
Seed selection, accelerate breeding process.
(3) the advantage allele that the present invention passes through the preferably molecular labeling, there is provided a kind of method of build seed selection, energy
Enough accelerate the genetic progress of duroc build (waistline character), the time of Duroc build improvement is shortened, so as to effectively carry
The economic benefit of high kind pig breeding, wherein, by the molecular labeling, the present invention is individual into CC types by the whole seed selections of TT types individual,
Then the overall waistline of Duroc colony will increase 3cm, improve build, improve meat yield.
Brief description of the drawings
Fig. 1 is that full-length genome of the duroc on waistline character on No. 4 chromosomes associates (GWAS) analysis chart；Its
In：Abscissa represents the chromosome numbers of pig；Ordinate expression-logP is worth；
Fig. 2 is the waistline correlation analysis figure of different genotype pig.
Embodiment
With reference to embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention are unlimited
In this.
Embodiment 1
(1) experimental animal
Experiment swinery colony is Du Luo for purebred U.S.A of Guangdong Wen Shi boars Science and Technology Ltd. used in the present embodiment
Grams 627, be boar branch company core group, colony's pedigree record is detailed.
American Duroc pig boar in the sources group, swinery free choice feeding, drinking-water, whole feeding side are chosen in this experiment altogether
Formula, rearing conditions etc. are always consistent, are conventional method.
Duroc litter size is less, and big group mean is only 9~10, but grows fast, feed conversion rate height, trunk lean meat
Rate is high, and intramuscular fat content is higher, strong stress resistance.More ripe corss combination system has been set up in the production of domestic goods pig.
Wherein the cross combination of DLY persistently keeps absolute superiority in the domestic and export markets in China.Because build is
Unique evaluation criteria of live body market price, and its abdominal circumference and waistline etc. are huge for pig type score influence, so Duroc
The research that Pigs Kidney encloses character is particularly important.
(2) sample is collected and phenotype records
Collect the docking of above-mentioned boar and ear tissue is soaked in the ethanol that volume fraction is 75% that to be placed in -20 DEG C of refrigerators standby
With.When pig is surveyed eventually, the waistline of pig is measured with tape, and have recorded 627 Duroc boar waistline data.
(3) pig full-length genome 50K SNP sentence type
The ear tissue or disconnected of each individual collection in the 627 Duroc herd boars chosen from above-mentioned sources group
Tail tissue, complete genome DNA is extracted with standard phenol-chloroform method, it is accurate through Nanodrop2000/2000C nucleic acid-proteins detector
Really DNA of the measure per a sample concentration and OD ratios (OD260/280, OD260/230).Through NanoDrop2000/2000C
Nucleic acid-protein detector detects qualified DNA sample, and DNA is diluted into 50ng/ μ L or so according to the concentration of detection.Again by 6 μ L
The DNA sample to be measured extracted mixes with 2 μ L Loading Buffer, is loaded to the Ago-Gel that mass fraction is 1%
In, electrophoresis 25min under 150V voltages, observe and take pictures under UV detector and gel imaging equipment, observe the complete of DNA
Whole property.
DNA sample Song Niuqin biotechnologies (Shanghai) Co., Ltd., the basis on Illumina Beadstration platforms
Company standard flow carries out pig full-length genome 50K SNP chips (Illumina, the U.S.) genotype and judged.Utilize R language
Checkmarker carries out quality control to all sample 50K chip scannings typing datas in GenABEL bags, rejects detection individual
Rate is less than 0.05 and Hardy-Weinberg equilibrium less than 90%, family Mendel error rate higher than 0.1, minimum gene frequency
Significance is higher than 10-6SNP, finally give 42642 SNP effective gene type data.
(4) full-length genome association (GWAS) analysis
In order to eliminate colony's stratification effect, the present invention is using the regression analysis of linear mixed model single-point and combines R language
GenABEL software kits carry out GWAS analyses, and the similarity correction stratification effect of genome between individual is utilized in analysis model.Using
Bonferrini methods determine SNP and waistline trait associations degree conspicuousness threshold value, and genomic level remarkable threshold removes for 0.05
With effective SNP site quantity, i.e. genome level of signifiance threshold value is 1.17 × 10-6, i.e., 0.05/42642 (effective SNP quantity)；
Chromosome level remarkable threshold is 1 divided by effective SNP site quantity, i.e. chromosome level of signifiance threshold value is 2.35 × 10-5, i.e., 1/
42642 (effective SNP quantity).
GWAS analysis results are as shown in Figure 1.From fig. 1, it can be seen that in Duroc, exist in No. 4 chromosome and significantly affect
The site that Pigs Kidney is enclosed, g.306T most strongly connected SNP is>C (P=7.70E-8)。
(5) correlation analysis of different genotype and waistline phenotype
It can be seen from table 1, the SNP site of molecular labeling is g.306T>C is extremely significantly correlated with waistline character, can be by right
The assisted Selection of this SNP site of duroc, so as to improve colony's waistline, and then accelerate breeding process.
Also known according further to table 1, TT types are lower than the average waistline of CC and CT types, illustrate that homozygote TT is to average waistline
It is worst.Further learnt by Fig. 2, homozygote TT and CC and CT genotype significant differences, and CC and TT genotype is even more
Difference is extremely notable, and this further illustrates that T allele is inferior position allele.Waistline is the important indicator of pig appearance traits, waist
Girth illustrates that the build of pig is sturdy.Therefore, more economic benefits can be brought by eliminating TT types and the pig of CT types, and we are being educated
The boar of superseded TT types and CT types is needed during kind, retains CC boar, to improve the allele C in the site by generation
Frequency.
The SNP site of the molecular labeling of table 1 is g.306T>C and waistline correlation
The target DNA sequence amplification of embodiment 2 and sequencing
(1) design of primers
Pass through Ensembl websites (http://asia.ensembl.org/index.html) download pig No. 4 chromosomes
Upper SEQ ID NO.1 DNA sequence dna.And design primer using primer-design software primer premier 6.0.Design is drawn
The DNA sequence dna of thing is as follows：
Primer C-F：5’-GGGTGAGGGTGGGATTAG-3’；
Primer C-R：5’-TTGCGTTAGTGATTGTCCTG-3’；
(2) PCR is expanded
The μ L of DNA profiling 1, μ L, the 2 × Tag PCR StanMixwith of distilled water 3.4 are added in 10 μ L reaction system
Each 0.3 μ L of Loading Dye 5 μ L, primer C-F and C-R.PCR reaction conditions are：After 94 DEG C of pre-degeneration 5min, 94 DEG C of denaturation
30s, 54.3 DEG C of annealing 30s, 72 DEG C of extension 45s, 35 circulations, last 72 DEG C of extensions 5min.
(3) determined dna sequence
Product after finally being expanded to PCR is sequenced, and sequencing is completed by Hua Da scientific ＆ technical corporation, genetic fragment sequencing
It is required that lead to for two-way survey.Sequencing result is as follows：
Note：The M marked in sequence table is mutational site, (is mutating alkali yl in bracket, is equipotential with display is underlined
Gene mutation), it is shown as designing primer sequence position in the head and the tail overstriking of the sequence.
The present invention is preliminary to carry out by being detected to the 306th bit base mutational site in SEQ ID NO.1 sequences
The application of association analysis between its genotype and the waistline character of pig, one is provided newly for the molecular marker assisted selection of pig
Molecular labeling.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
SEQUENCE LISTING
<110>Guangdong Wen'S Foodstuffs Group Co., Ltd.；Agricultural University Of South China
<120>A kind of molecular labeling for influenceing duroc waistline character and application
<130> 1
<160> 3
<170> PatentIn version 3.3
<210> 1
<211> 458
<212> DNA
<213> Artificial
<220>
<223>Influence the molecular labeling of duroc waistline character
<400> 1
gggtgagggt gggattagaa gtggtgccat gttggcgttc tctttgtaga aggcctagag 60
aggggatggc aggaagttat ggcttggaaa ttggatgagt gcggtggaaa caggtactaa 120
acggaaagca gaaagatctt ttatttactc ctttcttagt tgggcaacat ttctgggcca 180
cttgtcatgt tttcgtcagt gtactaagca tggagatagc cggatgaaca atgaagagtg 240
tatgcccttc aggagtttgc agatgaagga gcacacagac tcagacacca acgcctagag 300
cgtcgmgtga taagtgcttt ttgtggagat gcagagaaga gggctcctac cagagcttcc 360
tttgaacact gaattgaatc caggaggcgg tgcatacatt tgacatcttg tctcagaaat 420
gcaaaggtgg ctgaatacca ggacaatcac taacgcaa 458
<210> 2
<211> 18
<212> DNA
<213> Artificial
<220>
<223>Primer C-F
<400> 2
gggtgagggt gggattag 18
<210> 3
<211> 20
<212> DNA
<213> Artificial
<220>
<223>Primer C-R
<400> 3
ttgcgttagt gattgtcctg 20
Claims (10)
- A kind of 1. molecular labeling for influenceing duroc waistline character, it is characterised in that：Its SNP site refers to corresponding to international pig T on the edition 4 chromosome of genome 10.2 at 3400805bp>C is mutated.
- 2. the molecular labeling of the influence duroc waistline character described in claim 1 encloses character in identification Pigs Kidney and pig heredity is educated Applied in kind.
- A kind of 3. boar of the high lean meat percentage of molecular selection of influence duroc waistline character using described in claim 1 The method of strain, it is characterised in that comprise the following steps：Detect the molecular labeling of the influence duroc waistline character on No. 4 chromosomes of pig described in claim 1；Described molecule The SNP site of mark is T or C, eliminates T and retains C.
- 4. the boar according to claim 3 using the high lean meat percentage of molecular selection for influenceing duroc waistline character The method of strain, it is characterised in that：The boar is purebred American Duroc pig and its synthesis system.
- 5. a kind of primer pair for the molecular labeling for being used to identify the influence duroc waistline character described in claim 1, it is special Levy and be to include primer C-F and C-R, its nucleotide sequence is as follows：Primer C-F：5’-GGGTGAGGGTGGGATTAG-3’；Primer C-R：5’-TTGCGTTAGTGATTGTCCTG-3’.
- 6. the primer pair described in claim 5 influences to apply in duroc waistline character in identification.
- 7. the primer pair described in claim 5 is applied in duroc molecular marker assisted selection breeding.
- 8. the primer pair described in claim 5 is applied in the Duroc boar strain for cultivating high lean meat.
- 9. the method for the genetic improvement of a boar, it is characterised in that comprise the following steps：Determine the molecular labeling of the influence duroc waistline character in nucleus herds of breeding pigs described in the claim 1 of boar, and root According to the genotype in the above-mentioned site of pig corresponding selection is made during boar is chosen seeds：The Systematic Breeding world pig reference of boar CC types individual on the edition 4 chromosome of genome 10.2 at 3400805bp, eliminate the TT types and CT types individual of the point；With by In generation, improves the frequency of the allele C in the site, so as to the waistline of improved progeny pig.
- 10. the method for the genetic improvement of pig according to claim 9, it is characterised in that：Described boar includes Duroc and its synthesis system.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710852738.4A CN107586854B (en) | 2017-09-20 | 2017-09-20 | It is a kind of influence duroc waistline character molecular labeling and application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710852738.4A CN107586854B (en) | 2017-09-20 | 2017-09-20 | It is a kind of influence duroc waistline character molecular labeling and application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107586854A true CN107586854A (en) | 2018-01-16 |
| CN107586854B CN107586854B (en) | 2019-01-11 |
Family
ID=61047535
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710852738.4A Active CN107586854B (en) | 2017-09-20 | 2017-09-20 | It is a kind of influence duroc waistline character molecular labeling and application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107586854B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257529A (en) * | 2019-05-09 | 2019-09-20 | 华南农业大学 | SNP marker relevant to lean meat percentage, eye muscle area, eye muscle thickness and application on No. 6 chromosomes of pig |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106906303A (en) * | 2017-04-27 | 2017-06-30 | 江西农业大学 | One SNP marker for influenceing quality character of pork and its application |
| CN107177681A (en) * | 2017-06-08 | 2017-09-19 | 华南农业大学 | Application of the pig SNP marker in daily gain in pigs behavior study and/or pig breeding |
-
2017
- 2017-09-20 CN CN201710852738.4A patent/CN107586854B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106906303A (en) * | 2017-04-27 | 2017-06-30 | 江西农业大学 | One SNP marker for influenceing quality character of pork and its application |
| CN107177681A (en) * | 2017-06-08 | 2017-09-19 | 华南农业大学 | Application of the pig SNP marker in daily gain in pigs behavior study and/or pig breeding |
Non-Patent Citations (2)
| Title |
|---|
| PANT S D 等: "Comparative analysis of QTLs influencing obesity and metabolic phenotypes in pigs and humans", 《PLOS ONE》 * |
| YANG Q Y等: "Complex relationship between obesity and the fat mass and obesity locus", 《INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110257529A (en) * | 2019-05-09 | 2019-09-20 | 华南农业大学 | SNP marker relevant to lean meat percentage, eye muscle area, eye muscle thickness and application on No. 6 chromosomes of pig |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107586854B (en) | 2019-01-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108315433B (en) | It is a kind of influence Duroc boar intramuscular fat content molecular labeling and application | |
| CN110257529A (en) | SNP marker relevant to lean meat percentage, eye muscle area, eye muscle thickness and application on No. 6 chromosomes of pig | |
| CN108330197B (en) | One kind SNP marker relevant to Duroc kind fat thickness at back of pig and application thereof | |
| CN107365853B (en) | It is a kind of influence the effectively total teat number trait of pig molecular labeling and application | |
| CN108728552B (en) | It is a kind of influence duroc eye muscle area character molecular labeling and application | |
| CN110218798A (en) | SNP marker relevant to eye muscle area, eye muscle thickness and application on No. 7 chromosomes of pig | |
| CN110117665A (en) | SNP marker relevant to pig lean meat percentage and eye muscle area and application on No. 16 chromosomes of pig | |
| CN107164463A (en) | It is a kind of to be used for the SNP marker of measure and/or genetic improvement pig growth traits | |
| CN107177681A (en) | Application of the pig SNP marker in daily gain in pigs behavior study and/or pig breeding | |
| CN105624155B (en) | A kind of molecular labeling for influenceing pig feed conversion rate characteristic and application | |
| Teng et al. | Performance of whole genome prediction for growth traits in a crossbred chicken population | |
| CN108913779A (en) | A kind of SNP marker and its application influencing daily gain in pigs character | |
| CN108676899A (en) | A kind of and the relevant SNP marker of Erhualian sow litter trait and its detection method | |
| CN107828898A (en) | The SNP marker related to the kg body weight thickness of backfat of pig 100 and its application | |
| CN108559781A (en) | A method of cultivating high food utilization efficiency pig | |
| CN110144408A (en) | SNP marker relevant to total number of nipples and application on No. 7 chromosomes of pig | |
| CN110468212A (en) | SNP site relevant to fatty acid C14:0 content and application on No. 19 chromosomes of meat Simmental | |
| CN107267631A (en) | A kind of SNP marker for influenceing daily gain in pigs character and its application | |
| CN107858440B (en) | One kind SNP marker relevant to pig birth weight character and application thereof | |
| CN107988388B (en) | One kind SNP marker relevant to pig dewclaw undue growth character and application thereof | |
| CN113564264A (en) | SNP molecular marker located on No. 14 chromosome of pig and related to number of stillbirths and litter size of sow and application thereof | |
| CN112941198B (en) | SNP marker for detecting pig eye muscle area and application thereof | |
| CN107586854B (en) | It is a kind of influence duroc waistline character molecular labeling and application | |
| CN107779517B (en) | It is a kind of influence Duroc kind pig lean meat percentage character molecular labeling and its application | |
| CN108004332B (en) | It is a kind of to influence the molecular labeling and its application that the main hoof of pig is grown |
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 | ||
| CB02 | Change of applicant information |
Address after: 527400 9 East Causeway North Road, Xinxing County new town, Yunfu, Guangdong Applicant after: Winson food group Limited by Share Ltd Applicant after: South China Agricultural University Address before: 527400 9 East Causeway North Road, Xinxing County new town, Yunfu, Guangdong Applicant before: Guangdong Wens Foodstuff Group Co., Ltd. Applicant before: South China Agricultural University |
|
| CB02 | Change of applicant information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |