CN101906422B - LAGLS 9 gene as molecular marker relevant to birthweight characteristics of pigs as well as preparation method and application - Google Patents
LAGLS 9 gene as molecular marker relevant to birthweight characteristics of pigs as well as preparation method and application Download PDFInfo
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Abstract
The invention belongs to the technical field of preparation of molecular markers for livestock and particularly relates to a molecular marker and application, wherein the molecular is used for pig marker-assisted selection and is relevant to birthweight characteristics. The molecular marker is obtained by cloning an LAGLS 9 gene, the DNA sequence of the molecular marker is shown as a sequence table SEQ ID NO: 1. The basic-group mutation of A195-C195 is generated in the 195bp position of the sequence table SEQ ID NO: 1 so that the BtsC I-RFLP polymorphism is caused. The invention also discloses a primer for amplifying the partial sequences of the 3'-UTR zone of the LAGLS 9 gene and a polymorphism detection method and provides a new molecular mark for the pig marker-assisted selection.
Description
Technical field
The invention belongs to the molecular marker assisted selection technical field of pig, be specifically related to a kind of clone and application of the molecule marker relevant as the pig marker assisted selection with the birth weight proterties.Molecule marker of the present invention is relevant with the LGALS9 gene.
Background technology
Pig is important economic animal, and pork is because the tender delicious favor that extensively is subjected to the human consumer for a long time of its meat is one of main source of China people animal protein.In recent years, people grow with each passing day to the consumption of pork, how to improve production performance, one of focus into the pig breeding worker has reduced production costs.
Protocols in Molecular Biology develop the opportunity that for this reason provides important rapidly, rely on this technique means, scientists has been excavated out large quantities of and birth weight pig, the speed of growth, weaning weight, important reproductive trait such as litter size has the molecule marker of remarkable association.This provides theoretical foundation for the production performance that improves pig, and is greatly improved from making it in fact.
The birth weight of pig has indivisible close ties as an important reproductive trait with the postnatal speed of growth of pig and incubation rate.The influence of birth weight that recently a lot of bibliographical informations arranged to the speed of growth, and Li Jianhao (Li Jianhao. the landrace birth weight is to the influence of the speed of growth and nurture rate. piglet produces [J] .2006.18) discover that landrace 60 ages in days are heavy with 35 age in days weight averages and birth weight is proportionate and birth weight and the speed of growth are proportionate.(Beaulieu AD etc. such as Beaulieu, Impact of piglet birth weight, birth order and litter size on subsequentgrowth performance, carcass quality, muscle composition and eating quality of pork[J] .J Anim Sci.2010) discover, the growth speed of pigs that birth weight is lower is slower, thereby its time that comes into the market is increased.It is relevant that the birth weight that other there are some researches show piglet and incubation rate are also known, and birth weight piglet incubation rate when 1.0Kg is following rises along with the increase of birth weight; Birth weight piglet incubation rate when 1.0Kg is above does not have clear regularity.Birth weight during smaller or equal to 0.5Kg the piglet incubation rate only be 55.56% (the high foundation. painted face in Beijing opera pig birth weight is to the influence [J] of the speed of growth and incubation rate. herding and animal doctor, 1992,24 (1): 26).In addition, scientist finds the newborn infant that body weight is light more, its future trouble type ii diabetes of growing up, cardiovascular disorder and hypertensive risk are big more, one piece of research report in April in this year proves relevant (the Freathy RM etc. with the variation of gene of this phenomenon first, Variants in ADCY5 and near CCNL1 are associated with fetal growth and birthweight[J] .Nat Genet, 2010,42 (5): 430-435).Therefore, the clone of pig birth weight genes involved and evaluation be can be the genetic mechanism of explaining pig and other Mammals embryo growth and development important clue is provided, and provide theoretical foundation for the reproductive trait genetic improvement of pig.
Lectin semi-lactosi protein family be a class contain the carbohydrate identified region (carbohydrate recognition domain, CRD) can with β semi-lactosi bonded protein.Function comprise the extracellular function as with the mutual work of cell surface and extracellular matrix glycoprotein, glycolipid and endocellular function as with the mutual work of tenuigenin and nucleoprotein.Participate in regulation and control regulate several biological processes (Zick Y etc. such as cell adhesion, propagation, apoptosis, chemotactic, Inflammatory response and T necrocytosis, Role of galectin-8 as a modulator of cell adhesion and cell growth[J] .Glycoconj J, 2004,19:517-526; Lahm H etc., Tumor galectinology:insights into the complex network of a family ofendogenous lectins[J] .Glycoconj J, 2004,20:227-238; Friedrichs J etc., Contributions of galectin-3 and-9 toepithelial cell adhesion analysed by single cell force spectroscopy[J] .J Biol Chem, 2007,282:29375-29383).Semi-lactosi protein factor 9 (LGALS9) is a member of this protein family, belong to and repeat subfamily at random, contain a carbohydrate identified region respectively at aminoterminal and carboxyl terminal, thereby in conjunction with two kinds of different carbohydrates, two kinds of identified regions couple together (WadaI etc. by a joint albumen, Identification and characterization of galectin-9, a novel β-galactoside-binding mammalian lectin[J] .J BiolChem, 1997,272:6078-6086).Recent research shows, LGALS9 is that new uterine endometrium middle and advanced stage is secreted mutually and marker gene (the Popovici RM etc. of decidua, Galectin-9:a new endometrial epithelial marker for the mid-and late-secretory and decidualphases in humans[J] .J Clin Endocrinol Metab, 2005,90 (11): 6170-6176).Man month is in the decidua of cycle secretion phase uterine endometrium and early pregnancy, LGALS9 only expresses at epithelial cell, especially glandular epithelium and chamber epithelium expression amount are very high, and do not express (Shimizu Y etc. at stroma cell and immunocyte, Expression and localization of galectin-9 in the human uterodome[J] .Endocr J, 2008,55 (5): 879-887).Because the semi-lactosi protein family can combine with cell adhesion molecule such as ln and fibronectin, therefore the LGALS9 gene is considered to relevant with endometrial receptivity, may be middle albumen in conjunction with endometrial epithelial cell and blastomere, at the embryo is attached (the Gray CA etc. that may play a significant role in planting, Discovery and characterization of an epithelial-specific galectin in theendometrium that forms crystals in the trophectoderm[J] .Proc Natl Acad Sci USA, 2004,101:7982-7987).And there is certain relation the attached morning and evening of planting of embryo with birth weight, so this research intends inquiring into the relation of LGALS9 gene and birth weight.
Summary of the invention
The objective of the invention is to a kind of molecule marker relevant of clone from the LGALS9 gene, its preparation method and the application in the association analysis of pig marker assisted selection with the birth weight proterties as the pig marker assisted selection.
The present invention is achieved through the following technical solutions:
The applicant obtains part dna fragmentation with pig birth weight trait related gene from reporter gene LGALS9 clone, and its dna sequence dna is as described in sequence table SEQ ID NO:1 and the accompanying drawing 2.The base mutation of an A195-C195 is arranged at the 195bp place of sequence table SEQ ID NO:1, cause the BtscI-RFLP polymorphism.This base mutation is arranged in LGALS9 gene 3 '-UTR.
The preparation method of molecule marker of the present invention is:
With (the GenBank number of including: BV726873) be template design primer of pig STS sequence; Extract genomic dna, the design primer, the dna sequence dna of this primer is as follows: forward 5 ' GACATCCAGCTGACGCACG 3 ', reverse 5 ' GCTGGTGGCTTGGGACTAGG 3 '.
Pcr amplification, PCR product purification and order-checking obtain the nucleotide sequence shown in sequence table SEQ ID NO:1.
Sudden change detects the method for the PCR-RFLP that application is conventional to the 195th bit base shown in the sequence table SEQ ID NO:1, and tentatively carry out the application of the association analysis between its genotype and the pig birth weight proterties, for the molecular marker assisted selection of pig provides a new molecule marker.
More detailed technical scheme sees also " description of drawings " and the embodiment in " embodiment " of specification sheets.
Description of drawings
Sequence table SEQ ID N0:1 is the partial dna sequence of the pig immune trait genes involved LGALS9 that clones of the present invention, and the sequence total length is 375bp, this sequence the 195bp place base mutation of an A195-C195 is arranged.
Fig. 1: be the technology of the present invention schema.
Fig. 2: be the dna sequence dna of pig LGALS9 gene among the present invention, the mutational site shows with the overstriking italics that indicates underscore among the figure, and described primer sequence shows with italic overstriking and shade.
Fig. 3: be three kinds of genotype (AA AC CC) and the genome amplification electrophoresis result (P) of the BstcI-RFLP in pig LGALS9 gene 3 '-UTR district among the present invention, M:DNA molecular weight standard among the figure (DL50 ladder).
Fig. 4: be the A-C sudden change that the order-checking of pig LGALS9 gene forward is found among the present invention.
Embodiment
Embodiment 1: the amplification of pig LGALS9 Gene Partial dna sequence dna
(1) design of primers:
With the pig STS sequence of report (the GenBank number of including: be template BV726873), utilize biology primer-design software Primer Premier5.0, the increase primer of LGALS9 gene 3 '-UTR of design, the right dna sequence dna of this primer is as follows:
LGALS9: forward primer: 5 ' GACATCCAGCTGACGCACG 3 ', (corresponding to the 1-19 position of sequence table SEQ ID N0:1),
Reverse primer: 5 ' GCTGGTGGCTTGGGACTAGG 3 ' (corresponding to the 356-375 position of sequence table SEQ ID NO:1);
The dna sequence dna expanding fragment length of this primer is 375bp.
(2) pcr amplification reaction:
PCR reaction: the reaction cumulative volume is 10 μ l, pig DNA template 0.5 μ l wherein, distilled water 7.0 μ l, buffer 1 μ l, Mg
2+0.6 μ l, each 0.3 μ l of 10mM forward primer and reverse primer, dNTP 0.2 μ l, Taq enzyme 0.1 μ l (10U/ μ l).The PCR reaction conditions is: behind 94 ℃ of pre-sex change 5min, and circulate 35 94 ℃ of sex change 30s, 61 ℃ of annealing 30s, 72 ℃ of extension 25s, last 72 ℃ are extended 5min.The PCR product detects through 2% agarose gel electrophoresis.
(3) purifying of PCR product and order-checking
The purifying of PCR product: under ultraviolet lamp, contain the segmental gel of purpose from the cutting-out of low melting-point agarose gel, put into the 1.5ml centrifuge tube, use PCR product purification test kit (available from new the Changjiang river biotechnology (Beijing) company limited then, specification sheets operation according to this test kit) purified pcr product, concrete steps are the ratio adding GS damping fluids that add 400 μ l Gel-Shift binding buffer liquid (GS Buffer) according to the 100mg blob of viscose, put 50 ℃ of incubation 10min, the agarose blob of viscose is dissolved fully, put upside down mixing once in per two minutes; The glue that will dissolve is transferred to centrifugal adsorption column, and centrifugal adsorption column is placed the waste collection pipe, and the centrifugal 30s of 10000rpm discards waste liquid; Centrifugal adsorption column is put back in the waste collection pipe, added 500 μ l Wash Buffer in centrifugal adsorption column, the centrifugal 30s of 10000rpm abandons filtrate.Use 500 μ l Wash Buffer solution washings more once with same method; Centrifugal adsorption column is put in the meeting waste collection pipe the centrifugal 1min of top speed; The centrifugal adsorption column of careful taking-up is inserted in it in aseptic 1.5ml centrifuge tube, and central authorities add 30 μ l distilled waters at adsorption film, after room temperature leaves standstill 2-10min, and the centrifugal 1min of top speed; Take out centrifugal adsorption column, place-20 ℃ of preservations standby 1.5ml centrifuge tube (dna solution).
(4) determined dna sequence: sequencing is finished by Shenzhen Huada Genetic Technology Co., Ltd, and gene fragment is surveyed positive and negative two reactions.
Embodiment 2:PCR-RFLP diagnostic method is set up
(1) PCR-RFLP primer sequence (this primer also is the primer of amplification LGALS9 gene 3 '-UTR),
LGALS9 SNP: forward 5 ' GACATCCAGCTGACGCACG 3 ', (corresponding to the 1-19 position of sequence table SEQ ID NO:1), reverse 5 ' GCTGGTGGCTTGGGACTAGG 3 '.(corresponding to the 356-375 position of sequence table SEQ ID NO:1);
This primer amplification fragment length is 375bp.
(2) pcr amplification condition
PCR reaction cumulative volume 10 μ l, pig genomic dna template 1 μ l wherein, distilled water 7.1 μ l, 10 * damping fluid, 1 μ l, each 0.3 μ l of 10mM forward primer and reverse primer, dNTP 0.2 μ l, Taq enzyme 0.1 μ l (5U/ μ l).The PCR reaction conditions is: behind 94 ℃ of pre-sex change 5min, and circulate 35 94 ℃ of sex change 20s, 61 ℃ of annealing 30s, 72 ℃ of extension 20s, last 72 ℃ are extended 5min.The PCR product detects through 2% agarose gel electrophoresis.
(3) RFLP detects
PCR product endonuclease reaction volume is 10 μ l, PCR product 3 μ l wherein, distilled water 5.9 μ l, 10 * damping fluid, 1 μ l, restriction enzyme BstcI are 0.1 μ l (10U/ μ l), with centrifugal behind the sample mixing, 37 ℃ of incubators are placed 4h, detect enzyme with 4% agarose gel electrophoresis and cut the result, the record genotype is taken pictures under ultraviolet lamp.
Obtained 375bp specific amplification fragment with primer amplification pig genomic dna, sequencing result finds that this fragment finds an A-C transversion to be positioned at 3 '-UTR district (as shown in Figure 4) at the 195th, cause a BstcI restriction enzyme site (
), wherein the 197bp place is the polymorphism restriction enzyme site.Enzyme is cut and is produced three kinds of genotype, and the CC genotype has only 375bp one band, and the AA genotype has 197bp and 178bp two bands, and heterozygote AC genotype has 375bp, three bands of 197bp and 178bp, as described in Figure 3.
Embodiment 3: the polymorphism of molecule marker of the present invention in swinery used
Utilize PCR-BstcI-RFLP to detect " landrace " group (a kind of pig kind) on Guangdong Hua Nongwenshi herding limited-liability company water platform original seed pig farm with external pig blood relationship, the genotype frequency of swinery, gene frequency, shannon index (Longuet-Higgins MS.On the Shannon-Weaverindex of diversity, in relation to the distribution of species in bird censuses[J] .Theor PopulBiol, 1971,2 (3): 271-289) and χ
2Check P value is as shown in table 1, and the result shows that each genotype of LGALS9 gene all has distribution in the landrace kind, and AA genotype number is minimum, and AC genotype number is maximum, and landrace is that C allelotrope is preponderated; The shannon index of LGALS9 gene is higher, reaches 0.973, illustrates that landrace group's the diversity on this Guangdong Hua Nongwenshi herding limited-liability company water platform original seed pig farm is bigger; This gene is carried out the comptibility test of Hardy-Weinberg equilibrium law, and the result shows: the distribution of LGALS9 gene genotype and gene frequency meets Hardy-Weinberg equilibrium law (P＞0.05) in the landrace group.
The distribution of table 1 PCR-BstcI-RFLP polymorphism of the present invention in landrace
Embodiment 4: the application of molecule marker of the present invention in the association analysis of pig reproductive trait mark property
The present embodiment swinery is from the landrace group on Guangdong Hua Nongwenshi herding limited-liability company water platform original seed pig farm, and genotype detection has all been carried out in father and mother's generation and first filial generation, and body weight was measured in the filial generation birth in 0 day.
According to the group structure of collected specimens, the applicant use mixture model come statistical study LGALS9 gene SNP site the genotype effect and with the relation of birth weight proterties:
Y＝Xβ+Zγ+e
Wherein Y is a birth weight property determination value vector; X is the fixed effect incidence matrix; β is the fixed effect parameter vector, comprises the genotype effect of sex effect, parity effect, the other effect of line, candidate gene; Z is the incidence matrix of the stochastic effect in the mixture model; γ is all stochastic effect parameter vectors, comprises dam effect in male animal effect, the male animal; E is the random residual effect; The MIXEDMODELS program is carried out data processing and statistical study in employing SAS (Version 8.0) software.
Pig LGALS9 gene 3 '-UTR district BstcI-RFLP pleomorphism site and birth weight proterties are carried out association analysis, and as shown in Table 2: the AA genotype has 61 in 341 individualities, and the AC genotype has 153, and the CC genotype has 127.The result of the proterties significance of difference is as shown in table 2 between the different genotype, and the result shows that each genotype of pig LGALS9 gene 3 '-UTR district BstcI-RFLP pleomorphism site and landrace birth weight are extremely significantly related, and its P value is 0.0063.By the least square mean AA, AC and the genotypic individuality of CC are compared in twos, the result shows that the birth weight of AC genotype individuality is significantly higher than AA type and CC type individuality (the P value is respectively 0.0422 and 0.0026), but the individual birth weight of AA type and CC type does not have significant difference (p=0.6288).
The association analysis result of table 2 pig LGALS9 gene 3 '-UTR district A/C pleomorphism site genotype and birth weight
Annotate:
*Expression P＜0.01,
*Expression P＜0.05.
Claims (4)
1. molecule marker that detects with pig birth weight correlated character, its nucleotide sequence is as described in the sequence table SEQ ID NO:1, and the base mutation in that there is an A195-C195 at the 195bp place of sequence table SEQ ID NO:1 causes the BtscI-RFLP polymorphism.
2. a primer that detects molecule marker as claimed in claim 1 is right, and its nucleotide sequence is as follows:
Forward primer: 5 '-GACATCCAGCTGACGCACG-3 ',
Reverse primer: 5 '-GCTGGTGGCTTGGGACTAGG-3 '.
3. the application of the described molecule marker of claim 1 in the pig marker assisted selection.
4. the described primer of claim 2 is to the application in the pig marker assisted selection.
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| CN102776178B (en) * | 2011-05-12 | 2013-11-13 | 华中农业大学 | MicroRNA molecule marker associated with pig blood parameter properties, and application thereof |
| CN103289993B (en) * | 2012-03-01 | 2014-07-02 | 华中农业大学 | AP3D1 gene used as porcine birth weight character related molecular marker |
| CN103421768B (en) * | 2012-07-02 | 2014-11-19 | 华中农业大学 | Molecular marker related to piglet birth weight and use thereof |
| CN104630210B (en) * | 2013-11-13 | 2017-07-07 | 华中农业大学 | GLI2 genes are used as pig birth weight proterties related molecular marker and preparation method and application |
| CN107779516B (en) * | 2017-09-12 | 2018-10-19 | 华南农业大学 | It is a kind of influence pig birth weight character SNP marker and its application |
| CN112980962B (en) * | 2019-12-12 | 2022-05-31 | 深圳华大生命科学研究院 | SNP marker related to birth weight trait of pig and application thereof |
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Non-Patent Citations (4)
| Title |
|---|
| Uenishi.H,et al.AK235094.1.<<GeneBank>>.2007,第1页. * |
| 兰旅涛等.3个外来种猪群ESR基因位点多态性及其与繁殖性能相关性分析.《江西农业大学学报》.2006,(第01期), * |
| 岳敏等.西藏小型猪MSTN基因5'调控区的酶切多态性分析.《实验动物与比较医学》.2008,(第03期), * |
| 张令钢等.美系大白猪雌激素受体基因的多态性分析.《上海交通大学学报(农业科学版)》.2008,(第04期), * |
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