CN105063037A - Hu sheep prolactin gene SNP locus and application thereof - Google Patents
Hu sheep prolactin gene SNP locus and application thereof Download PDFInfo
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- CN105063037A CN105063037A CN201510524900.0A CN201510524900A CN105063037A CN 105063037 A CN105063037 A CN 105063037A CN 201510524900 A CN201510524900 A CN 201510524900A CN 105063037 A CN105063037 A CN 105063037A
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Abstract
The invention discloses a Hu sheep prolactin gene SNP locus and an application thereof. The SNP locus includes a PRL4591 locus, a PRL1228 locus, a PR2351 locus and a PR4591 locus. By studying the Hu sheep prolactin gene SNP locus, it is of remarkable significance to increasing the lactation yield or improving the milk character of Hu sheep.
Description
Technical field
The present invention relates to sheep luteotropin gene SNP site and application thereof, belong to technical field of molecular biology.
Background technology
Sheep originates in Taihu Lake basin, has that maternal instinct is good, a feature of the strong and early sexual maturity of milk performance.Again because of its how litter-bearing proterties, the improved seeds of domestic mutton sheep crossbreeding and improvement are now become.In recent years, mainly concentrate on the polymorphism research to its economic characters functional gene to the research of sheep genetic breeding, the correlative study for Milk Production is relatively less.But along with this sheep kind fecund polyembryony, nutrition relative deficiency, ewe Milking ability are low, thus cause lamb development bad, lower for the resistance, and survival rate declines, and make the advantage of polyembryony fecund fail to play.Therefore this test intended is started with from Molecular and genetic basis, and research prolificacy sheep lactation gene, to finding the marker gene controlling sheep lactation amount size, for sheep molecular breeding provides technical support.
Research shows that the chemical element in different types of sheep Ruzhong is substantially identical, but its milk yield is subject to the impact of the factors such as heredity, parity, environment (V.M.Russo2013), age, trophic level, sunshine, band cub number, physiological status, milking processes.The milk character of domestic animal is a quantitative character, and by controlled by multiple genes, the research in recent years for lactation gene mainly concentrates on ox.The major function of luteotropin gene (PRL) be promote the mammogenesis of mammalian subject, milk generates, start and play a significant role in maintenance lactation.Therefore, PRL gene is regarded as candidate gene domestic animal milk performance to vital role.Sheep prolactin (PRL) full length gene is 8468bp, containing 5 exons and 4 introns.
Milk character is a kind of quantitative character typically controlled by multiple minor gene, is the less proterties of sheep Recent study.At present the domestic and international weave construction to luteotropin gene, the mechanism of action and and the relation of the reproductive trait such as lactation amount still know little about it, also needing to do a large amount of exploration work to these several respects, providing a new approach to the high-efficient breeding for improving sheep reproductivity.Therefore, how obtaining the SNP site of sheep luteotropin gene, to improve sheep lactation amount or sheep milk character, is this area technical problem urgently to be resolved hurrily.
Summary of the invention
Technical problem to be solved by this invention is to provide sheep luteotropin gene SNP site and application thereof.To achieve these goals, the present invention adopts following technical proposals:
The present invention relates to sheep luteotropin gene SNP site, it is characterized in that described SNP site is positioned in PRL gene order.
In a preferred embodiment of the present invention, described SNP site is PRL4591 site, PRL1228 site, PR2351 site and/or PR4591 site.
In a preferred embodiment of the present invention, it is characterized in that described PRL4591 site is CC type.
The invention still further relates to a kind of sheep luteotropin gene, it is characterized in that described gene PRL4591 site is CC type, PRL1228 site is AC type, PR1039 site is AA type, PR2351 site is AA type or GG type and/or PR4591 site are CC type.
The present invention also relates to the application of above-mentioned sheep luteotropin gene SNP site in sheep breeding on the other hand.
In a preferred embodiment of the present invention, described application is in the application improving lactation amount or improve in sheep milk character.
The present invention, by research sheep luteotropin gene SNP site, has significant meaning for improving lactation amount or improving sheep milk character.
Embodiment
Below in conjunction with specific embodiment, technology contents of the present invention is described in detail.
1 materials and methods
1.1 experimental animal phenotypic datas are collected and blood specimen collection
The sheep 41 lactation period that silver ring sheep stud random choose parity (1-2), age, body condition, the expected date of childbirth, nutritional status etc. are basically identical beyond the Great Wall in Midong District, Urumchi.Laboratory animal is mainly fed pulse family and gramineous grass, concentrates and supplements a fine fodder.Flock of sheep equal stable breeding, free choice feeding and drinking-water.Vein takes blood sample, heparin sodium anti-freezing ,-20 DEG C of preservations.
The collection of breast and the mensuration of milk-content: the 4th day (allowing lamb as far as possible eat up colostrum) after production 20 point, by lamb and ewe isolation after, adopt the mode of artificial lactation to squeeze milk, morning 7 next day point, 13: 30,20 lactation amounts measuring ewe respectively.Measure and day send laboratory by early, middle and late for the milk sample of collection rear-20 DEG C of freezen protective of 25ml mixing of respectively getting.
Adopt the FOSS5000 series of tests instrument of Denmark's import to detect sample, wherein IntegratemilkTestingTMFossmatic5000 is for measuring milk-content, and IntegratedMilkTestingTMMilkoScanFT4000 is used for measuring somatic number.Testing index comprises protein ratio (%), milk fat content (%), milk sugar rate (%), total solids etc.
The extraction of 1.2 genomic dnas
Phenol-chloroform method is adopted to extract the genomic dna of sheep, sterilizing ,-20 DEG C of preservations after TE dissolved dilution.Detect DNA purity with spectrophotometer, be then diluted to 50ng/ μ l for subsequent use.
1.3PCR primer and reagent
According to the sheep luteotropin gene sequence (GenBank accession number NW_004080183.1) that GenBank announces, utilize primer-design software Primer5.0 to design primer (table 1), synthesized by the raw work in Shanghai; Proteinase K, taq enzyme are all purchased from Takara company, and phenol, chloroform, ethanol are all purchased from Jinan chemical reagents corporation.
PCR reaction system and reaction conditions
PCR reaction system: 10 × buffer is (containing Mg
2+) 5 μ L, each 2.5 μ L of 10pmol upstream and downstream primer, Taq DNA polymerase 3U, dNTPs (2.5umol/L) 5 μ L, DNA1 μ L, adds ultrapure water to 50 μ L.
Adopt grads PCR to increase to the primer, determine the suitableeest annealing temperature of often pair of primer.The reaction conditions of PCR: 95 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 52-62 DEG C of annealing 45s, 72 DEG C extend 1-2min, altogether 35 circulations; 72 DEG C extend 5min.4 DEG C of preservations.The product of pcr amplification is detected with 2% agarose gel electrophoresis.
1.5 statistical study
BioEdit, Mega6.0 and Chromas is adopted to obtain the sequence of PRL gene to order-checking institute and peak figure compares, detection SNP site.By SPSS13.0 statistical software statistic data, fixed model is utilized to analyze PRL genotype to the hereditary effect of milk production trait.Test of significance is carried out to the Traits change of surveyed sheep different genotype.
Table 1PRL full genome primer
1.6PRL gene PCR result
Adopt the annealing temperature of grads PCR to primer to screen, select PCR primer 1.5% agarose gel electrophoresis and detects, result display, band is clear, and without primer dimer, specificity is good, after serve Hai Shenggong and check order.
The examination of 1.7SNP site
The PRL gene sequence data using the raw work in Bioedit arrangement Shanghai to return, compares with MEGA6.0.Use Chromas to check peak figure, find out 23 SNP site.
Find after single for PRL gene SNP site genotype is carried out association analysis with sheep milk character lactation period: only PR4591 site is relevant with lactation amount, other 4 sites are all relevant to milk character.Result is as table 2-6.
The remarkable analysis of table 2 sheep PR4244 site and lactation amount
The correlation analysis of the single SNP site of table 3 sheep and milk character
The correlation analysis of table 3 sheep luteotropin gene PR1228 site and milk character
The correlation analysis of table 4 sheep luteotropin gene PR1039 site and milk character
The correlation analysis of table 5 sheep luteotropin gene position PRL2351 point and milk character
The correlation analysis of table 6 sheep luteotropin gene position PR4591 point and milk character
As can be seen from table 2,3,4,5,6, it is relevant to SNP site that sheep has 4 growth traitss, shows that SNP site has statistical significance.PRL4591 site CC type is significantly higher than AA type (P<0.05) at the 8th week lactation amount; AC type later stage milk fat content pole, PRL1228 site is significantly higher than CC type (P<0.01), and mid-term, total solids AC type was significantly higher than CC type (P<0.05); Milk fat content pole in AA type early stage, PR1039 site is significantly higher than AC type and CC type; PR2351 site AA type and GG type milk fat content in mid-term are significantly higher than AG type (P<0.05), AA type albumen in mid-term rate pole is significantly higher than AG type and GG type, and AA type and AG type pole are significantly higher than GG type (P<0.01); CC type early stage milk fat content pole in PR4591 site is significantly higher than AA type (P<0.01), and late protein rate and total solids CC type in mid-term pole are significantly higher than AA type (P<0.01).
The above is the preferred embodiments of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from principle of the present invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.For the technician in general field, to any apparent change that it does under the prerequisite not deviating from connotation of the present invention, all by formation to infringement of patent right of the present invention, corresponding legal obligation will be born.
Claims (6)
1. sheep luteotropin gene SNP site, is characterized in that described SNP site is positioned in PRL gene order.
2. SNP site according to claim 1, described SNP site is PRL4591 site, PRL1228 site, PR2351 site and/or PR4591 site.
3. SNP site according to claim 1, is characterized in that described PRL4591 site is CC type.
4. a sheep luteotropin gene, is characterized in that described gene PRL4591 site is CC type, PRL1228 site is AC type, PR1039 site is AA type, PR2351 site is AA type or GG type and/or PR4591 site are CC type.
5. the application of above-mentioned sheep luteotropin gene SNP site in sheep breeding.
6. application according to claim 5, described application is in the application improving lactation amount or improve in sheep milk character.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110387373A (en) * | 2019-07-24 | 2019-10-29 | 上海交通大学 | Chinese ring-necked pheasant stimulating milk secretion fibroin and its encoding gene and application |
| CN110878362A (en) * | 2019-12-18 | 2020-03-13 | 南昌师范学院 | Detection method for correlation between PRL gene 5' regulatory locus point and chicken testicular character and application |
| CN113265476A (en) * | 2021-07-21 | 2021-08-17 | 中国农业大学 | Gene chip, molecular probe combination, kit and application for analyzing milk production performance of sheep |
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| CN101063169A (en) * | 2007-06-04 | 2007-10-31 | 西北农林科技大学 | PCR-RFLP method for detecting goat prolactin gene single nucleotide polymorphism |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110387373A (en) * | 2019-07-24 | 2019-10-29 | 上海交通大学 | Chinese ring-necked pheasant stimulating milk secretion fibroin and its encoding gene and application |
| CN110878362A (en) * | 2019-12-18 | 2020-03-13 | 南昌师范学院 | Detection method for correlation between PRL gene 5' regulatory locus point and chicken testicular character and application |
| CN110878362B (en) * | 2019-12-18 | 2023-03-28 | 南昌师范学院 | Detection method for correlation between PRL gene 5' regulatory locus point and chicken testicular character and application |
| CN113265476A (en) * | 2021-07-21 | 2021-08-17 | 中国农业大学 | Gene chip, molecular probe combination, kit and application for analyzing milk production performance of sheep |
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