内容简介:  

黄　菁　王少丽　乔传令
33
(中国科学院动物研究所,农业虫鼠害综合治理研究国家重点实验室　北京　100080)
Automated programming of degenerate primers and the cloning of the diamondback esterase gene. HUANG
Jing , WANG Shao2Li , QIAO Chuan2Ling
33 ( Institute of Zoology , Chinese Academy of Sciences , the State Key Labo2
ratory of Integrated Management of Pest Insects and Rodents , Beijing 　100080 , China) .
Abstract 　It is a normal strategy to clone protein family cDNA using the degeneracy of the code to design PCR prim ers from protein multiple2sequence alignments. With the tools of blastp , blockmaker , CodeHop , SwissProt , and SpTrEMBL , degenerate primers were designed for cloning of the insect esterase cDNA. Using one pair of primers , a partial cDNA was obtained from the diamondback moth by RT2PCR. Similarity alignment showed that the products of the cloned cDNA were very similar to those of insect esterases. The results indicated that these degenerate primers de2 signed by this software could be used to obtaining specific RT2PCR product.
Key words 　degenerate primer design , CodeHop insect esterase gene , RT2PCR
摘　要　利用遗传密码简并性,针对特定的氨基酸序列设计简并引物,是克隆蛋白质家族cDNA 的常规方法。文章介绍了利blastp ,blockmaker ,CodeHop ,SwissProt ,SpTrEMBL 等网络工具及数据库设计昆虫抗性酯酶的简并引物。用这对引物从抗有机磷杀虫剂的小菜蛾Plutella xylostella 中克隆了1 段cDNA。经blastx 检索genebank ,发现此cDNA 产物与其它昆虫抗性酯酶基因有高度的相似性。研究表明,程序化设计的简并引物可信性强,阳性率高,能迅速得到满意结果。
关键词　简并引物设计, CodeHop , 昆虫酯酶基因, RT2PCR

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