草鱼羧肽酶A4基因部分序列多态性及生长性状关联分析

【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...

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Published in南方农业学报 Vol. 43; no. 3; pp. 380 - 384
Main Author 曹婷婷 刘小献 白俊杰 于凌云
Format Journal Article
LanguageChinese
Published 上海海洋大学水产与生命学院,上海201306%大连海洋大学生命科学与技术学院,辽宁大连,116023%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 2012
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
Subjects
SNP位点
关联分析
基因型
生长性状
羧肽酶A4基因
草鱼
基因型
羧肽酶A4基因
SNP位点
关联分析
草鱼
生长性状
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ISSN2095-1191
DOI10.3969/j:issn.2095-1191.2012.03.380

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Summary:【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。
Bibliography:45-1381/S
grass carp; CPA4 gene; SNP sites; genotype; growth traits; correlation analysis
[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 a
ISSN:2095-1191
DOI:10.3969/j:issn.2095-1191.2012.03.380