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凡纳滨对虾微卫星DNA的筛选及其特性的研究
The isolation and characterization of microsatellite markers from Litopenaeus vannamei
【摘要】 采用小片段克隆法构建凡纳滨对虾的部分基因文库,用放射性同位素[γ-32P]ATP标记的(CA)15、(AT)12、(AG)12、(AAT)8、(AAG)8做探针筛选阳性克隆。共获得微卫星序列152个,其中二核苷酸为核心的微卫星序列:(AG)n>(AC)n>(AT)n,三核苷酸为核心的微卫星序列:(AAT)n>(CAT)n>(AAG)n。应用引物设计软件primer3.0设计引物105对。选择合成10对理论上易出现影子带的引物,筛选后用31个凡纳滨对虾个体对这些引物进行了评估,结果10对引物中有8对引物能扩增出谱带,其中1对扩增产物出现影子带,1对扩增产物为单态,有6对扩增产物出现多态。
【Abstract】 The partial genome library of Litopenaeus vannamei was constructed by method of small fragments DNA cloning and the positive clones were screened with the probes (CA) 15 , (AT) 12, (AG) 12, (AAT)8 and (AAG)8 labeled with [γ- 32P] ATP. 152 sequences containing microsatellites were obtained. The dinucleotide repeats were (AG)n>(AC)n>(AT)n and the trinucleotide repeats were (AAT)n>(CAT)n>(AAG)n respectively. 105 sets of primers were designed with the software primer 3.0. 10 primers that the stutter bands easily occur in theory were synthesized and 31 samples were tested with them. Eight loci can give bands and six had a degree of polymorphisms, one was monomorphic and one had stutter bands.
【Key words】 Litopenaeus vannamei; microsatellite DNA; repeat sequences; clone;
- 【文献出处】 上海水产大学学报 ,Journal of Shanchai Fisheries University , 编辑部邮箱 ,2007年01期
- 【分类号】S917.4
- 【被引频次】13
- 【下载频次】293