中国对虾微卫星DNA的筛选  

ISOLATING MICROSATELLITE DNA OF CHINESE SHRIMP PENAEUS CHINENSIS

作　　者：徐鹏[1] 周岭华[1] 相建海[1] 

机构地区：[1]中国科学院海洋研究所,青岛266071

出　　处：《海洋与湖沼》2001年第3期255-259,共5页Oceanologia Et Limnologia Sinica

基　　金：国家重点基础研究发展计划项目!资助 ;G1 9990 1 2 0 0 7号;国家自然科学基金!资助项目 ;3 9970 1 1 3号

摘　　要：于 2 0 0 0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 1 8个阳性重组克隆中 ,其中perfect共2 3个 ,占 74% ,imperfect2个 ,占 7% ,compoundperfect 1个 ,占 3% ,compoundimperfect 5个 ,占 1 6%。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。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℃ 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.

关 键 词：中国对虾 微卫星 筛选 DNA 基因 阳性克隆 

分 类 号：Q959.223[生物学—动物学]