中华绒螯蟹一个种质鉴定相关RAPD标记的SCAR转化  

SCAR Transformation of a RAPD Marker for Germplasm Identification in Chinese Mitten Crab Eriocheir sinensis

作　　者：许志强[1,2] 葛家春[1,2] 李跃华[1] 朱清顺[1] 潘建林[1] 

机构地区：[1]江苏省淡水水产研究所,江苏南京210017 [2]南京师范大学生命科学学院,江苏南京210097

出　　处：《水产科学》2012年第7期429-432,共4页Fisheries Science

基　　金：江苏省科技支撑计划项目(BE2009336);江苏省农业科技自主创新资金资助项目(cx(10)125);江苏省水产三项工程项目(K2008-2)

摘　　要：通过克隆测序获取了中华绒螯蟹Opp17目的片段,分析其长度、碱基组成等序列特征。序列测定结果表明,克隆片段的准确大小为1170bp,G+C含量为50.34%,Opp17(TGACCCGCCT)引物与所测序列中的引物结合位点完全匹配。在GenBank中对序列进行BLast检索,未找到任何同源序列。在此基础上,在原有随机引物Opp17的基础上向3'端延伸13～19个nt以设计SCAR引物,最终设计了1对中华绒螯蟹特异性SCAR(sequence characterized ampLified regions,序列特征化扩增区)引物SCAR-1(F-TGACCCGCCTCAGCACCCGAACG;R-TGACCCGCCTCTCAGCCACACACC)。利用这对引物对长江天然群体中华绒螯蟹基因组DNA进行PCR扩增,结果表明所有个体均能扩增出预期大小1170bp的特异性条带。对目的条带进行回收测序,结果与之前的测序结果完全一致,表明本研究已成功获取了中华绒螯蟹种质鉴定国家标准中Opp17目的条带的SCAR标记。The molecular biology marker（a specific RAPD band） of the national standard for germplasm identification in Chinese mitten crab Eriocheir sinensis（GB/T 19783-2005） was determined by cloning sequencing.The length of fragment was 1170 bp,with average G＋C contents of 50.34%,which contained the binding sites of the random primer Opp17（TGACCCGCCT）.The sequence was subjected to do BLAST search against nucleotide databases,and found no homology to known nucleotide sequences in GenBank.After gel extraction,cloning and sequencing of the specific RAPD bands,a pair of SCAR（sequence characterized amplified regions） primers named SCAR-1 were designed according to the sequence information（F-TGACCCGCCTCAGCACCC GAACG;R-TGACCCGCCTCTCAGCCACACACC）.PCR amplification with SCAR-1 was carried out in the natural population of the crab to determine the specificity of amplification of the primers.The results showed that a specific band about 1000 bp was amplified in the whole 23 samples.The fragment was cloned and sequenced again to confirm the specificity of the primers,indicating that a specific SCAR molecular biology marker for the germplasm identification in the crab（GB/T19783-2005） was successfully developed in the study.

关 键 词：中华绒螯蟹 种质鉴定 SCAR 

分 类 号：Q78[生物学—分子生物学]