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中国对虾微卫星DNA的筛选
徐鹏, 周岭华, 相建海
中国科学院海洋研究所 青岛266071
摘要:
于2000年2月以中国科学院海洋研究所水族楼暂养的中国对虾为材料，采用常规方法从尾部肌肉中提取DNA，构建小片段部分基因组文库。采用人工设计合成的(CT) 7、(AT) 7重复片段作引物，利用PCR法对中国对虾小片段部分基因组文库进行筛选。本实验首次在中国对虾中获得31个微卫星序列，分别分布于18个阳性重组克隆中，其中perfect共23个，占74%，imperfect2个，占7%，compound perfect 1个，占3%，compound imperfect5个，占16%。结果还表明，在中国对虾基因组DNA中，(CT) n、(AT) n形式的微卫星序列的含量非常丰富。
关键词:  中国对虾  微卫星  筛选
DOI：
分类号:
基金项目:国家重点基础研究发展计划项目资助，G1999012007号;国家自然科学基金资助项目，39970113号
ISOLATING MICROSATELLITE DNA OF CHINESE SHRIMP PENAEUS CHINENSIS
XU Peng, ZHOU Ling Hua, XIANG Jian Hai
Institute of Oceanology,The Chinese Academy of Sciences,Qingdao,266071
Abstract:
Thirty-one microsatellites of the Chinese shrimp Penaeus chinensis were obtained through PCR screening small size fractionated libraries of P. chinensis with simple tandem repeats primers (AT) n and (CT) n. The bacterial suspensions were used in PCR. A high temperature of 95°C was used for breaking the bacteria. After screening, recombinant positive clones containing microsatellites were sequenced. Among the 31 microsatellites in 18 positive recombinant positive clones are 23 perfect ones (74%), 2 imperfect (7%), 1 compound perfect (3%), and 5 compound imperfect (16%). The experiment shows that microsatellite sequences characterized by (AT) n and (CT) n abundant in genomic DNA of Chinese shrimp. Primers can be designed according to these microsatellite franking sequences and used in genetic analysis.
Key words:  Penaeus chinensis, Microsatellite, Screening
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