CN101580880B - 评价牛个体的胴体重量的基因标记及使用其的胴体重量评价方法 - Google Patents
评价牛个体的胴体重量的基因标记及使用其的胴体重量评价方法 Download PDFInfo
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- 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
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- C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
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- C—CHEMISTRY; METALLURGY
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Abstract
本发明提供使用基因标记的评价牛个体的胴体重量的评价方法。确定牛NCAPG基因的e9位点的碱基,该碱基为G时,评价为胴体重量大。或者,确定牛NCAPG蛋白的E9位点的氨基酸,该氨基酸为甲硫氨酸时,评价为胴体重量大。
Description
技术领域
本发明涉及评价牛个体的胴体重量(carcass weight)的基因标记及使用该基因标记的胴体重量评价方法。
背景技术
牛的肉质和胴体重量是直接关系到价格的经济性状,对于怎样评价关于这些性状的遗传能力来帮助牛的改良的问题,一直以来设计并采用基于育种值的方法等。
肉质和胴体重量被认为是涉及多个基因的量的性状。如果可以确定对肉质和胴体重量有较大影响的基因或基因组区域(QTL),从而进行优良的基因型的判别,则可以将其用于牛的改良。
目前为止,报道了根据采用黑毛和种种公牛的父系半同胞家系的QTL分析,在牛6号染色体上存在影响体重或胴体重量的基因组区域(Takasuga等(2007)Mamm.Genome 18卷125-136页)。然后,在另外的黑毛和种种公牛中,在6号染色体的相同区域中确认到QTL(Setoguchi等(2006)日本动物遗传育种学会第7届大会摘要(Abstract of the 7th meeting of Japanese Society of Animal Breeding and Genetics))。另一方面,在某些褐毛和种种公牛及继承其优良的遗传性状的子代种公牛中也在与上述基本相同的区域检出了QTL。
发明内容
然而,实际上并不清楚是怎样的遗传信息承载了优良的遗传性状,因此在评价牛个体的胴体重量时,无法使用基因型等遗传信息。
于是,本发明的目的在于提供使用基因标记的评价牛个体的胴体重量的评价方法。
本发明人通过详细分析牛6号染色体上的影响体重或胴体重量的基因组区域,发现在NCAPG基因的SNP中,e9位点的SNP是6号染色体上的体重或胴体重量QTL的关键SNP或与关键SNP存在连锁不平衡的SNP,藉此,包括NCAPG基因的e9位点且e9位点的碱基为G的DNA可用作使胴体重量增加的基因标记,还发现e9位点的碱基为G的SNP为显性突变以及具有该SNP的NCAPG基因编码E9位点的氨基酸为甲硫氨酸的突变NCAPG蛋白等情况,从而完成了本发明。
于是,本发明的评价牛个体的胴体重量的评价方法的特征在于,确定NCAPG基因的e9位点的碱基或NCAPG蛋白的E9位点的氨基酸。
此外,本发明的牛NCAPG基因的e9位点为G。本发明的牛NCAPG蛋白的E9位点的氨基酸为甲硫氨酸。
此外,本发明的DNA具有包括牛NCAPG基因的e9位点的所述基因的部分或全部,该e9位点的碱基为G。
此外,本发明的评价牛个体的胴体重量的基因标记由具有包括牛NCAPG基因的e9位点的所述基因的部分或全部的DNA构成。
此外,本发明的选择胴体重量大的牛个体的选择方法包括:对于各牛个体,确定NCAPG基因的SNP中的e9位点的碱基的步骤;选择在NCAPG基因的至少一个等位基因中该碱基为G的个体的步骤。
此外,本发明的使野生型牛个体的胴体重量增加的方法的特征在于,在不进行交配的情况下,使用基因重组技术,制造在NCAPG基因的至少一个等位基因中将e9位点的碱基置换为G的牛,或者制造表达E9位点的氨基酸为甲硫氨酸的NCAPG蛋白的牛。
此外,本发明的牛具有编码E9位点的氨基酸为甲硫氨酸的NCAPG蛋白的外源性DNA。该外源性DNA可以是表达该NCAPG蛋白的表达载体。
如果采用本发明,则可以提供使用基因标记的评价牛个体的胴体重量的评价方法。
具体实施方式
以下,参照实施例,对基于上述发现而完成的本发明的实施方式进行详细说明。实施方式和实施例中没有特别说明的情况下,使用记载于以下标准试验方案集中的方法,或者对其进行改进或修改后的方法:J.Sambrook,E.F.Fritsch和T.Maniatis编,《分子克隆实验指南(Molecular cloning,a laboratory manual)》(第三版),冷泉港出版社(Cold Spring Harbor Press),冷泉港(Cold Spring Harbor),纽约(2001);F.M.Ausubel,R.Brent,R.E.Kingston,D.D.Moore,J.G.Seidman,J.A.Smith,K.Struhl编,《新编分子生物学实验指南(Current Protocols in Molecular Biology)》,约翰威立出版有限公司(John Wiley&Sons Ltd.)等。此外,在使用市售的试剂盒和测定装置的情况下,没有特别的说明时,则使用它们所附的试验方案。
本发明的目的、特征、优点和其思路通过本说明书的记载,从业者可以了解,根据本说明书的记载,只要是从业者就可以容易地再现本发明。以下记载的发明的实施方式和具体的实施例等是本发明的优选实施方式,是为了示例或说明而例举的,本发明并不局限于这些实施方式。在本说明书中所揭示的本发明的意图及范围内,基于本说明书的记载,可以进行各种改进和修改,这一点对于从业者是可以了解的。
牛NCAPG基因的SNP
牛野生型NCAPG基因的e9位点的碱基为T,但如实施例所示,牛NCAPG基因的e9位点的碱基为G时,胴体重量增加。因此,如果确定NCAPG基因的SNP中的e9位点的碱基,则可以评价或预测胴体的重量。
在这里,e9位点是指序列编号1的牛NCAPG基因的cDNA(NM_001102376)中的第1372个碱基以及牛基因组中的NCAPG和转录产物hnRNA、NCAPG基因同源物等中与该碱基对应的所有碱基。
牛野生型NCAPG蛋白的E9位点的氨基酸为异亮氨酸,但e9位点的碱基为G的牛突变NCAPG基因编码E9位点的氨基酸为甲硫氨酸的突变NCAPG蛋白。因此,可以确定牛NCAPG蛋白的E9位点的氨基酸来代替NCAPG基因的e9位点的碱基。
在这里,E9位点是指序列编号2的牛NCAPG蛋白(NP_001095846)中的第442个氨基酸以及部分肽或NCAPG同源物等中与该氨基酸对应的所有氨基酸。
基因标记
本发明中,评价牛个体的胴体重量时的诊断标记是指用于检测牛NCAPG基因的e9位点的SNP的基因相关物质。例如,包括含NCAPG基因的DNA、作为转录物的hnRNA和mRNA、作为翻译物的多肽、作为最终产物的蛋白质等。
诊断标记为NCAPG基因等DNA的情况下,为了检测上述SNP,能够确定具有SNP的碱基即可。具体来说,可以直接确定碱基序列,也可以采用PCR或RFLP,对检测方法没有特别限定。诊断标记为作为NCAPG基因的转录产物的hnRNA或mRNA的情况下,可以通过确定RNA序列来检测SNP。直接检测这些SNP的情况下,测序的核酸不需要包括整个NCAPG基因,可以是NCAPG基因或cDNA的一部分,包括具有SNP的碱基(这里为e9位点的碱基)且能够确定该碱基即可。
诊断标记为NCAPG蛋白等肽的情况下,为了检测上述突变,可以通过常规方法直接确定具有突变的氨基酸。直接检测该突变的情况下,测序的肽不需要包括整个NCAPG蛋白,可以是NCAPG蛋白的一部分,包括具有突变的氨基酸(这里为E9位点的氨基酸)且能够确定该氨基酸即可。
SNP的判定方法
e9位点的碱基的种类以分子生物学方式确定即可,例如从牛细胞提取基因组DNA,通过常规方法确定e9位点的碱基。如果该e9位点的碱基为G的杂合型或G的纯合型,则可以作出该牛的胴体重量大的评价。
E9位点的氨基酸也例如使用抗体等从牛细胞中纯化NCAPG蛋白,通过常规方法确定氨基酸序列即可。如果该E9位点的氨基酸为甲硫氨酸,则可以作出该牛的胴体重量大的评价。
此外,使用该评价方法,可以从大量的牛中选择胴体重量大的牛个体。即,对于各牛个体,确定NCAPG基因的SNP中的e9位点的碱基,选择在至少一个等位基因中该碱基为G的个体,或者确定NCAPG蛋白的E9位点的氨基酸,选择具有至少该氨基酸为甲硫氨酸的突变NCAPG蛋白的个体,从而可以选择胴体重量大的牛个体。
在这里,NCAPG基因在牛中是高度保守的,因此作为实施本发明的对象的牛的种类为黑毛和种、褐毛和种、荷斯坦种等,没有特别限定。
SNP的人工操作
因在NCAPG基因的至少一个等位基因中e9位点的碱基为G而表达E9位点的氨基酸为甲硫氨酸的NCAPG蛋白的突变牛中,如实施例所示,胴体重量增加。该NCAPG基因中,是否有e9位点以外的突变与胴体重量的增加无关。
因此,为了使野生型牛个体的胴体重量增加,在不进行交配的情况下,使用敲除动物制造、敲减动物制造、转基因动物制造等众所周知的个体的基因重组技术,制造在NCAPG基因的至少一个等位基因中将e9位点的碱基置换为G的牛,或者制造表达E9位点的氨基酸为甲硫氨酸的NCAPG蛋白的牛即可。
目前为止,使用牛获得了胚胎干细胞(Biochem.Biophys.Res.Commun.309卷104-113页2003),也制成了敲除牛(Nat Ganet 36卷671-672页2004)。使用这样的发育工程学的基因重组技术,在牛个体中将特定的碱基置换为目标碱基也是可能的。
于是,为了使在NCAPG基因的两个等位基因中都不具有G作为e9位点的碱基的牛个体的胴体重量增加,例如制造在NCAPG基因的至少一个等位基因中将e9位点的碱基置换为G的牛即可。该情况下,该突变等位基因是显性的,因此并不需要置换两个等位基因,仅置换一方即可。
或者,如实施例所示,由于该突变是显性突变,因此通过制造牛个体中表达NCAPG蛋白的E9位点的氨基酸为甲硫氨酸的突变蛋白的牛,也可以制造胴体重量增加的牛个体。具体来说,例如制造导入有表达该突变蛋白的表达载体的转基因牛即可。
实施例
以下,使用实施例进行更详细的说明。
[1]DNA的提取以及微卫星多态和SNP的分型
基因组DNA从精液、肾周围脂肪或血液通过常规方法提取。使用可以特异性地扩增目标基因组片段的引物,通过PCR法扩增该基因组区域。对于微卫星多态,荧光标记反向侧引物,将PCR扩增产物用ABI 3730 DNA分析仪(应用生物系统公司(ァプラィドバィォシステム社))进行电泳后,通过GENESCAN和GeneMapper软件(应用生物系统公司)分析,从而进行分型。对于SNP,使用Big Dye Terminator v.3.1循环测序试剂盒(应用生物系统公司)进行PCR扩增产物的直接测序,从而确定序列,进行SNP的检测和分型。对于表2的SNP 19,因为重复序列是多型的,所以荧光标记反向侧引物,将PCR扩增产物用ABI 3730 DNA分析仪(应用生物系统公司)进行电泳后,通过GENESCAN和GeneMapper软件(应用生物系统公司)分析,从而进行分型。
[2]胴体重量的测定方法
胴体重量使用屠宰场出售的牛的胴体的分级成绩。
[3]与胴体重量相关的SNP的统计学处理
本实施例中,NCAPG基因的e9位点产生的向G的突变是显性突变,显示对胴体重量产生影响。
对于在牛6号染色体上检出了胴体重量或体重QTL的3头黑毛和种种公牛(A~C)和2头褐毛和种种公牛(D、E)的基因组DNA,通过使用牛基因组序列制成的多种微卫星标记和SNP标记进行分型比较。这时,为了分清通过各标记得到的2个等位基因的基因型是来源于相同染色体的优良型(Q)还是非优良型(q),对于各种公牛的子代也进行了分型。表1中表示所用的引物。
【表1】
cM | 标记名 | 正向引物(序列编号) | 反向引物(序列编号) | 碱基 |
DIK9014 | AGCCAGCTQAGTCAAATTCC(3) | GTGAGACAGATGGCCAATCA(4) | 37,780,130 | |
45.93 | DIK4852 | TCAGCTTCTGTACCCATGGAC(5) | AGCCAGCGTTTCCAGAAAAG(5) | 37,855,588 |
SNP 0 | CACCATGTCCTGACCTCAQAT(53) | TAACAGTGCCCTGCATGAGQA(54) | 38,009,208 | |
DIK9015 | CCTTTGTTTGCTGGGTCAAT(7) | GGGCTTGATCTCTGGTTGAG(8) | 38,051,344 | |
DIK9016 | ATGGCAACCCACTACTCCAG(9) | TTGCTACCAAGCAAGCACTG(10) | 38,162,665 | |
DIK9017 | GTAAACTCAAGCCACGGCA(11) | CGACAACCTTGATGTGACAAA(12) | 38,670,448 | |
DIK9018 | GATGGCACTGQAGGTAGAGC(13) | CAACCCCATGGATTGTAACC(14) | 38,948,770 |
cM:连锁地图上的位置(Ihara等(2004)Genome Res.14 1987-1998)
碱基:以牛基因组序列(2007-9-13)中的引物的第一个碱基的编号表示的牛6号染色体上的位置(http://www.hgsc.bcm.tmc.cdu/)
其中,SNP 0的碱基表示SNP的位置
其结果表明,包括NCAPG基因的约660kb(SNP0-DIK9017)的区域在5头种公牛的优良型等位基因中共通,且5头种公牛都包含可以与非优良型等位基因区别开的标记。
对存在于该区域的4个基因的蛋白质翻译区域中存在的SNP进行了检索,发现了5处在种公牛A中为杂合且伴随氨基酸置换的SNP。针对它们,考察了各种公牛,结果5头种公牛都以杂合具有的SNP仅有e9位点的SNP。
对于包括该SNP的邻近的19个SNP(表2),考察了对胴体重量的影响。
【表2】
位置(碱基):以牛基因组序列(2007-9-13)中的碱基的编号表示的牛6号染色体上的位置(http://www.hgsc.bcm.tmc.cdu/)
MAF:190头黑毛和种中的次要等位基因频率(Minor allele frequency)GenBank登录号:LOC523874 XM_602183;NCAPG NM_001102376;LOC540095 XM_001250262在这里,用于PCR的引物示于表3。
【表3】
SNP ID | 正向序列(序列编号) | 反向序列(序列编号) |
SNP 1 | TGTACCTTGTGATACATGCTTTAAAAT(15) | GATCTGTGCACAATAGGAGTTCAATAA(18) |
SNP 2 | CACAGGGGAGTTGAATAGCAG(17) | CCTGTTGCTTCCAAGTAGACC(18) |
SNP 3 | CAGAAGCAGCTGACACAGGA(19) | ACTCACAGACTGCTGCATCG(20) |
SNP 4 | GGAGAAAACCCACAAGCTCA(21) | GCCTCCGAGACAAAGTTTCA(22) |
SNP 5 | GGGATGTTGGCAGAAAAGAA(23) | CATCCCAAATATTTTTCAAAGG(24) |
SNP 6 | TTGTAGATAATTTTCTTAGGTGAAGGA(25) | GGACACTCTTTCCTAAACCTTTT(26) |
SNP 7 | TTCTCACTTAATGGGGAGCTG(27) | TTAGGAGAGCAAATTAGAACAAGAG(28) |
SNP 8 | TTTCAGAATGTGAATTTTGGCTTA(29) | AGCCAAAAGCACTGAAAACAC(30) |
SNP 9 | TTTCAGAATGTGAATTTTGGCTTA(31) | AGCCAAAAGCACTGAAAACAC(32) |
SNP 10 | TGGATACTGTTTGGAGTTTTGTG(33) | TCAGTCGGGCACATACAGAA(34) |
SNP 11 | TGGATACTGTTTGGAGTTTTGTG(35) | TCAGTCGGGCACATACAGAA(38) |
SNP 12 | TTCTGTATGTGCCCGACTGA(37) | TCTGGCAGCTAAATTAAGCAAA(38) |
SNP 13 | TTTACTTTTGGTGGGGGATG(39) | TGCTAAAAATGACCTTGCACA(40) |
SNP 14 | GAGCTTACATGGGGAGGGTTA(41) | CTTCAAGAAATGAGCACCAAA(42) |
SNP 15 | AGTATTTGGTGCTCATTTCTTGA(43) | TGAATTTAATTAGAAAAACTCTTCCAT(44) |
SNP 16 | GCTGCTTTTGGGACTGATTG(45) | GCAGCAGCAAGACATTGAAA(45) |
SNP 17 | TTTTAAGCTCAATGGAATCAGGA(47) | TGGAATCGCACACCAGAAAT(48) |
SNP 18 | ATGGGGTACCTCACAGCACT(49) | AAGAAAACCTGAATCTTTTTCACC(50) |
SNP 19 | CGCCGCTCGTATGTAAATG(51) | TGAACTGACCCGAAAGGAAG(52) |
首先,对7990头黑毛和种阉牛的胴体重量上端群(570-670kg;上端4.7%)中的94头(同一种公牛的子代至多5头)、下端群(290-410kg;下端4.6%)中的96头(同一种公牛的子代至多5头)进行分型,以2×2表进行了Fisher的精确检验(参照表3“p值”),与该e9位点的相关性最高(表4的SNP 9:p(等位基因数)=1.2×10-11)。
【表4】
SNP ID | p值(等位基因数) | p值(显性模型中的检验) | p值(隐性模型中的检验) |
SNP 1 | 9.9E-05 | 2.1E-04 | 0.011 |
SNP 2 | 1.0E-04 | 5.1E-06 | 0.0072 |
SNP 3 | 4.4E-05 | 3.1E-04 | 0.0035 |
SNP 4 | 0.0010 | 0.0030 | 0.091 |
SNP 5 | 1.0 | ND | ND |
SNP 6 | 1.0 | ND | ND |
SNP 7 | 0.91 | ND | ND |
SNP 8 | 0.82 | ND | ND |
SNP 9 | 1.2E-11 | 6.7E-11 | 0.012 |
SNP 10 | 0.0037 | 0.0032 | 0.20 |
SNP 11 | 0.012 | 0.0066 | 0.16 |
SNP 12 | 0.0057 | 0.0041 | 0.12 |
SNP 13 | 1.0 | ND | ND |
SNP 14 | 0.0016 | 0.0030 | 0.091 |
SNP 15 | 0.82 | ND | ND |
SNP 16 | 1.6E-10 | 6.1E-10 | 0.024 |
SNP 17 | 0.83 | ND | ND |
SNP 18 | 0.0081 | 0.0056 | 0.12 |
SNP 19 | 0.0081 | 0.0056 | 0.12 |
ND:种公牛A为纯合等位基因,所以未进行检验。
接着,使用fastPHASE程序(Scheet,P.和M.Stephens(2006)Am J HumGenet 78,629-644.),推测由19个SNP构成的单倍型,结果仅该e9位点为G的单倍型在胴体重量上段群内的频率比下段群内的频率大(表5的单倍型5和6:对于这些单倍型和除此之外的单倍型的2×2表的Fisher的精确检验值为p=6.7×10-11)。
【表5】
由此可知,对胴体重量产生影响的突变是在NCAPG基因的e9位点产生的G,该突变是显性突变。
[4]作为标记的利用
对种公牛A-D的子代进行分型,考察与胴体重量的相关性。结果示于表6和表7。
【表6】
种公牛D(母子代)以外的子代都为阉牛
【表7】
贡献率:基因型所说明的效果在表现型值的总方差中所占的比例种公牛D(母子代)以外的子代都为阉牛
对于以NCAPC基因的e9位点的向G的突变判定的胴体的重量增加效果,一个等位基因的突变(杂合个体)通常显示出效果,即使两个等位基因发生突变(纯合个体),与杂合个体相比,虽然效果根据家系而不同,但效果也比最初的突变小。由此确认,该突变是不完全显性的。
此外,不仅是特定的家系,对任意群的375头进行分型的结果中也得到同等的结果,由此可知,该SNP可以广泛地被用作可以判别使胴体重量增加的基因型的优良的标记。
序列表
<110>社团法人畜产技术协会
<120>评价牛个体的胴体重量的基因标记及使用其的胴体重量评价方法
<130>P1547
<160>54
<170>PatentIn version 3.1
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ggtaggcgaa cgtgaacagg ctttgtctcg gccgggtact ggcgccatgg ggaaggagaa 60
gagactgctg ctgattaagg aggccttcca gctggcgcag cagcctcacc agaaccaggc 120
gaagctggtg gtggcgctga accgcaccta cggctcggtg gatgacaaaa cagattttca 180
tgaggagttt gttcattacc ttaaatatgc tatggtggtc tataaacgag aaccagctgt 240
ggaaagagta atagaatttg ccgcaaagtt tgttacttca tttcaccaat cagatatgga 300
aaatgatgaa gaggaggagg aggatggtgg cattttaaat tatttgctta cttttctatt 360
aaagtctcat gaagcaaaca gcaatgcagt tagatttaga gcgtgccagc tcataaacaa 420
gctcttggga aatatgccag aaaatgccca aattgatgat gatttgtttg ataaaattaa 480
tgaagccatg cttattagat tgaaagataa agttccaaat gtaaggatac aggcagttct 540
tgctctttca cgccttcagg atcccaaaga tgatgaatgc ccagtggtta atgcatatgc 600
tactttgatt gaaaatgatt caaatccaga agttaggcgg gcagtgttat cgtgtattgc 660
gccatcagca aagactttgc caaaaattgt tgggcgcacc aaggatgtga aagaaactgt 720
cagaaagctg gcttatcagg ttttagctga aaaggttcac atgagagctc tgtccattgc 780
tcagagagta atgctccttc aacaaggtct caatgaccga tcagatgctg tgaaacaagc 840
aatgcagaag catcttctcc aaggctggtt acgttttact gaaggaaata tattagagtt 900
gcttcatcga ttggatgtgg aaaattcttc tgaagtagca gtctctgttc tcaatgcctt 960
gttttccatg actcctctta atgaactggc agaaatctgt aaaaataatg acggcaggaa 1020
attgattcca gcagatacat taactcctga atttgctttg tattggcgtg tcctttgtga 1080
acatttgaaa tcaaaaggag aagaaggtga agaattttta gagcagattt tgccagagcc 1140
tgtagtatat gcagagtatt tactgagtta tattcaaagc attccagttg ttactgaaga 1200
acagagaggt gatttttcct atattggcaa tttgatgaca aaagaattca taggtcaaca 1260
attaattcta attatcaagt ctttggatac caatgaagaa ggaggaagga aacgaatact 1320
gggtatctta caggagattc ttactctacc taccacacca atatccctaa tttcttttct 1380
tgttgagaga ctgctccaca tcattataga tgataataag agaatacaaa ttgttacaga 1440
aattatctca gagattcggg cacccattgt tactgttgct gttaataatg atccagctga 1500
tgcaagaaag aaagagctta agatggccga aataaaagtt aaacttattg aggcaaaaga 1560
ctctttggaa aattgcatta ccttacagga ttttcatcga gcatcagaat taaaagaaga 1620
aataaaagca ttagaggatg ccaaaataaa ccttttgaaa gagacagagc aacatgaaat 1680
gaaagaagtc cacatagaga agaatgatgc tgaaacccta cagaagtgtc ttattttatg 1740
ctatgaacta ttgaagcaga tgtccacttc aacaggtata ggtgcaacca tggatggcat 1800
cattgaatct ttgattcttc ctggaataat aaatgttcat cctgtagtaa gaaatttggc 1860
tgtactgtgt ttgggatgct gtggactgca gaatcaggat tttgcaagta aacactttgt 1920
attactcttg caggttttgc aaattgatga tgtgacaata aaaataagtg ctttaaaggc 1980
aatctttgac caactgatga catttggatt tgaaccattt aaaactaaaa aaatcaaagc 2040
tactcaaaag gaaggtgcag aaataaactc cagtgaagag caagagtcaa aagaatccga 2100
agaagagaca gctatagcca agaatgttct gaaactactt tccgatttct tagatagtga 2160
ggtgtctgaa ctcagaacag gagctgcaga aggactagcc aagctgatgt tctctggact 2220
tttggtcagc agcaggattc tttctcatct tgtcttgtta tggtacaacc ctgtgactga 2280
agaggacatt cgacttcgac attgcctcgg cgtgttcttc cccatgtttg cttatgcaag 2340
caggactaac caggaatgtt ttgaagaagc ctttcttcca actctgcaaa cactggccaa 2400
tgcccctgcg tcatctcctc tagctgaaat agatataact aatgttgctg agttacttgt 2460
agatttgaca agaccaagtg ggttaaatcc tcaggccaag aatcccccag attatcaggc 2520
cttaacagtt catgacaatc tggctatgaa aatttgcaat gagatcctaa catgtccaca 2580
ttcaccagaa gttcgggtct atacgaaagc tttgagttct ttagaactca gcagcgatct 2640
tgctaaagat cttctggttg tgctgaatga gattctggag caagtaaaag atagaacatg 2700
tctaagagct ctggagaaaa tcaagattca gatagaaaaa ggaattaaag aacatagtga 2760
ccaagctgta gcagcacagg atgacatcac aactatgact gttcttcaga gtgaagatga 2820
aaagaataaa gatgtataca taactcctgt caaggaagta aaagcaactc gaatgaaatc 2880
cactcagcaa aagaccaaca gaggacggag aaaagtggta gcttcagcta gaacgaacag 2940
aagatgtcag actattgaag ctgaggctaa ctctgaaagt gatcatgaag ttccagaacc 3000
agaatcagaa atgaagatga gattaccaag acgagccaaa acagcagcac tagaaaaaag 3060
taaacttaac cttgcacaat ttctcaatga agatacaagt taggagaaga aatgatggag 3120
gtggagtcct ttgaaaaatg gcctttaaaa ttatgttcag ttctttgctt taataaagtt 3180
acccttgtat gaaaattaaa gtctgattct tgcagaaaaa aaaaaaaaaa a 3231
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<211>20
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ttctgtatgt gcccgactga 20
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tctggcagct aaattaagca aa 22
<210>39
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tttacttttg gtgggggatg 20
<210>40
<211>21
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tgctaaaaat gaccttgcac a 21
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<211>21
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gagcttacat ggggagggtt a 21
<210>42
<211>21
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cttcaagaaa tgagcaccaa a 21
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agtatttggt gctcatttct tga 23
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tgaatttaat tagaaaaact cttccat 27
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gctgcttttg ggactgattg 20
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gcagcagcaa gacattgaaa 20
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ttttaagctc aatggaatca gga 23
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atggggtacc tcacagcact 20
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aagaaaacct gaatcttttt cacc 24
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cgccgctcgt atgtaaatg 19
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<211>20
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tgaactgacc cgaaaggaag 20
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<211>21
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caccatgtcc tgacctcaga t 21
<210>54
<211>20
<212>DNA
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taacagtgcc ctgcatgaga 20
Claims (7)
1.一种评价牛个体的胴体重量的方法,其特征在于,确定NCAPG基因的e9位点的碱基或NCAPG蛋白的E9位点的氨基酸,
所述碱基为G或所述氨基酸为甲硫氨酸时,评价为使个体的胴体重量增加的遗传能力高于所述碱基为T的牛个体,
所述e9位点是指序列编号1的牛NCAPG基因的cDNA(NM_001102376)中的第1372个碱基,
所述E9位点是指序列编号2的牛NCAPG蛋白(NP_001095846)中的第442个氨基酸。
2.一种牛NCAPG基因,其特征在于,e9位点的碱基为G,所述e9位点是指序列编号1的牛NCAPG基因的cDNA(NM_001102376)中的第1372个碱基。
3.一种牛NCAPG蛋白,其特征在于,E9位点的氨基酸为甲硫氨酸,所述E9位点是指序列编号2的牛NCAPG蛋白(NP_001095846)中的第442个氨基酸。
4.一种DNA,其特征在于,具有包括牛NCAPG基因的e9位点的所述基因的部分或全部,所述e9位点的碱基为G,所述e9位点是指序列编号1的牛NCAPG基因的cDNA(NM_001102376)中的第1372个碱基。
5.一种评价牛个体的胴体重量的基因标记,其特征在于,由具有包括牛NCAPG基因的e9位点的所述基因的部分或全部的DNA构成,所述e9位点是指序列编号1的牛NCAPG基因的cDNA(NM_001102376)中的第1372个碱基。
6.一种选择胴体重量大的牛个体的方法,其特征在于,包括:
对于各牛个体,确定NCAPG基因的e9位点的碱基的步骤;
选择在NCAPG基因的至少一个等位基因中的所述e9位点的碱基为G的个体的步骤,
所述e9位点是指序列编号1的牛NCAPG基因的cDNA(NM_001102376)中的第1372个碱基。
7.表达载体,其特征在于,表达E9位点的氨基酸为甲硫氨酸的NCAPG蛋白,所述载体具有编码所述E9位点的氨基酸为甲硫氨酸的NCAPG蛋白的DNA,所述E9位点是指序列编号2的牛NCAPG蛋白(NP_001095846)中的第442个氨基酸。
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CN103866001A (zh) * | 2014-01-17 | 2014-06-18 | 甘肃农业大学 | 牦牛胴体性状候选基因检测试剂盒及其检测方法 |
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