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王春艳, 俞小牧, 童金苟. 鲤鱼多态性EST标记的筛选与特性分析[J]. 水生生物学报, 2008, 32(2): 207-212.
引用本文: 王春艳, 俞小牧, 童金苟. 鲤鱼多态性EST标记的筛选与特性分析[J]. 水生生物学报, 2008, 32(2): 207-212.
WANG Chun-Yan, YU Xiao-Mu, TONG Jin-Gou. DEVELOPMENT AND CHARACTERIZATION OF POLYMORPHIC EST MARKERS IN COMMON CARP[J]. ACTA HYDROBIOLOGICA SINICA, 2008, 32(2): 207-212.
Citation: WANG Chun-Yan, YU Xiao-Mu, TONG Jin-Gou. DEVELOPMENT AND CHARACTERIZATION OF POLYMORPHIC EST MARKERS IN COMMON CARP[J]. ACTA HYDROBIOLOGICA SINICA, 2008, 32(2): 207-212.

鲤鱼多态性EST标记的筛选与特性分析

DEVELOPMENT AND CHARACTERIZATION OF POLYMORPHIC EST MARKERS IN COMMON CARP

  • 摘要: 表达序列标签(Expressed sequence tag,EST)标记在基因组作图和分子标记辅助育种研究中具有重要价值。筛选和开发可用于遗传作图的鲤鱼多态性EST标记,对研究鲤鱼的基因组结构、遗传多样性研究和遗传育种有重要意义。本研究根据GenBank数据库中鲤鱼EST序列设计了67对EST引物,有47对在鲤鱼基因组DNA中成功扩增得到稳定的特异性条带,经单链构象多态性(SSCP)分析,12对(25.5%)引物扩增的EST在1个鲤鱼回交家系中具有多态性,其中6个为鲤鱼功能基因,5个与斑马鱼(Danio rerio)功能基因具较高相似性,1个与斑点叉尾(Ictaluruspunctatus)功能基因具较高相似性。这些多态性的EST在鲤鱼二倍体家系和单倍体群体中的等位基因分离均符合孟德尔规律(1:1或3:1),单倍体中的SSCP条带数目为二倍体的二分之一。选用其中4对多态性的EST引物对洞庭湖鲤鱼进行初步群体遗传分析,结果显示基因多样性(H)为0.437,远低于微卫星标记所揭示的该群体基因多样性(接近于1)。结果表明,多态性的EST-SSCP标记尽管遗传变异性较低,但是作为来源于编码区的Ⅰ型遗传标记,在遗传作图和种群遗传适应性等研究中有较好的应用潜力。

     

    Abstract: Expressed sequence tags (ESTs) are of high value in genome mapping and marker-assisted selection (MAS).Thisstudy aims to develop EST markers for genome mapping and other studies of common carp (Cyprinus carpio L).A total of 7381EST sequences (400.3500bp) were downloaded from the EST database of theGenBank as the source data of the present analy-ses.Sixty-seven pairs of primers (designated as HB1-67) were designed in the 3? untranslated region (UTR) of the common carpESTs,47 of which can successfully amplify specific products fromgenomicDNA.Twelve polymorphicEST markers were identifiedin a backcross mapping family of common carp by single-strand conformation polymorphism (SSCP) analysis.Six of the 12 poly-morphicEST markerswere derived from common carp genes of known function, and the rest were from common carp ESTs homolo-gous to putative genes in zebrafish (Danio rerio) or channel catfish (Ictaluruspunctatus).Alleledistribution of these polymorphicEST markers was found to follow theMendelian segregation (1.1 or 3.1) in both the backcross family and gynogenetic haploids,and the number of SSCP bands in haploid population was half of that in diploid family.Thegene diversity (H=0.437) of Dongt-ing Lake population of common carp revealed by 4 polymorphic EST markers is much lower than that by previous microsatelliteanalysis (close to 1).The results of present study indicated that EST-SSCP markers,with codominant inheritance, can be used asa powerful tool in the genetics of not only diploid but also haploid common carp.As TypeI marker derived from coding region withrelatively lower genetic variability, EST-SSCP markers are still of high potentials in such studies as genome mapping (includinggenetic linkagemaps and genemaps, as well as comparative mapping), genetic structure, and genetic adaptability of fish popula-tions.

     

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