草鱼羧肽酶A4基因部分序列多态性及生长性状关联分析  

Polymorphism amongst several CPA4 gene sequences and correlation analysis of growth traits in grass carp

作　　者：曹婷婷[1,2] 刘小献[3] 白俊杰[1] 于凌云[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所/农业部热带亚热带水产资源利用与养殖重点实验室,广州510380 [2]上海海洋大学水产与生命学院,上海201306 [3]大连海洋大学生命科学与技术学院,辽宁大连116023

出　　处：《南方农业学报》2012年第3期380-384,共5页Journal of Southern Agriculture

基　　金：国家高技术研究发展计划项目(2011AA100403);现代农业产业技术体系建设专项资金项目(Nycytx-49-03)

摘　　要：【目的】筛选出更多与草鱼生长相关的SNP位点标记,为草鱼分子育种研究提供理论依据。【方法】根据草鱼EST数据库中属于消化酶的羧肽酶A4基因的cDNA序列设计引物,采用PCR产物直接测序法,对含预计SNP位点的cDNA序列进行扩增、测序,并寻找新的SNP位点。【结果】cDNA上预计的SNP位点不存在,但在内含子3上检测到两个SNP位点,分别位于内含子的第40、241个碱基处,命名为A40G位点和G241T位点。经创造酶切位点的限制性片段长度多态检测(CRS-RFLP),发现A40G位点AA型占50.3%、AG型占23.8%、GG型占25.9%,G241T位点的GG型占48.6%、GT型占47.9%、TT型占3.5%。利用一般线性模型(GLM)将两个SNP位点与草鱼6个生长性状进行关联分析,结果表明,A40G位点的GG型个体在体重、体长、体高均高于AA型和AG型,但差异不显著(P>0.05);G241T位点TT型个体最重,GG型最轻,但差异不显著(P>0.05);A40G和G241T两个位点组成的7种双倍型个体的体重、体长、体高、尾柄高等重要生长性状差异也不显著(P>0.05)。【结论】在草鱼羧肽酶A4基因的内含子3上发现两个SNP位点(A40G位点和G241T位点),每个SNP位点都可分为3种基因型,但两个SNP位点的不同基因型以及由其组成的双倍型与草鱼的重要生长指标均不存在显著相关。[Objective]Many SNP sites regarding the growth of grass carp were screened to provide a theoretical reference for grass carp molecular breeding.[Method]Primers were designed from CPA4 gene cDNA sequences belonging to digestive enzymes from EST database of grass carp.cDNA sequences,expected to contain SNP sites,were amplified and sequenced by direct sequencing of PCR product,and new SNP sites were also targeted.[Result]It was found that expected SNP sites didn＇t exist.Two nucleotide mutations were identified in 40th and 241th basic group of intron 3 by using direct sequencing and blasting,which were named as 40G and G241T.The polymorphisms were genotyped by CRS-RFLP methods.The percentage of AA,AG,and GG genotype in A40G site was recorded as 50.3,23.8 and 25.9%,respectively.The percentage of GG,GT,and TT genotype in G241T was recorded as 48.6,47.9,and 3.5%,respectively.A general linear model（GLM） was used to statistically analyze the relationship between two SNP sites and major growth traits.The result indicated that in A40G site individuals from GG genotype were higher than AA and AG genotype in terms of body weight,length and height,however,there was no significant difference（P0.05）.In G241T site,individuals from TT genotype were the heaviest,and the ones from GG genotype were the lightest,but there was no significant difference（P0.05）.There was no significant difference in body weight,length and height etc.of 7 douple-type individuals from A40G and G241T.[Conclusion]Two SNP sites found in intron 3（A40G G241T） from the CPA4 gene in grass carp were divided into three genotypes,nevertheless,various genotypes of the two SNP sites and their double types were not significantly associated with essential growth indices of grass carp.

关 键 词：草鱼 羧肽酶A4基因 SNP位点 基因型 生长性状 关联分析 

分 类 号：S965.112[农业科学—水产养殖]