CONSTRUCTION OF A cDNA LIBRARY OF HOST RECOGNITION KAIROMONE FOR TELENOMUS THEOPHILAE  

野蚕黑卵蜂寄主识别利它素的cDNA文库构建(英文)

作　　者：高其康[1] 胡萃[1] 等 

机构地区：[1]浙江大学应用昆虫学研究所,杭州310029

出　　处：《Entomologia Sinica》2002年第1期35-39,共5页中国昆虫科学（英文版）

基　　金：国家自然科学基金 (项目号 30 0 70 5 15 );浙江省自然科学基金的资助

摘　　要：Based on the kairomone from the accessory gland of female Bombyx mori elicting the parasitoid, Telenomus theophilae, to parasitize artifical beads. A cDNA expression library of enrich kairomone gene was constructed using mRNA rich secretory portion of the accessory glands of B. mori at the 9 day old of pupa. The titer of the unamplified library was 3.29×10 4 pfu/μL with a recombinant rate of 90.05%, and titer of amplified library was 1.56×10 7 pfu/μL. The inserted cDNA fragments of the library ranged from 0.5 to 8.0 kb and concentrated around the 2.5 kb, 1.1kb, and 0.75kb regions. All of the data suggest that the library has appropriate titer and high frequency kairomone gene fragments by using special developing stage and tissue of B. mori to extract mRNA. Directional cloning and λZiplox expression were help to clone the kairomone gene with immunoscreening.根据家蚕性附腺中含有的利它素能诱导野蚕黑卵蜂寄生涂有该利它素的人造卵及能用家蚕卵繁育野蚕黑卵蜂的特性 ,用化蛹 9天的家蚕雌蛹性附腺分泌部的高丰度的mRNA构建了富含利它素基因的cDNA文库。构建成的未扩增文库的滴度为 3.2 9× 10 4 pfu μL ,重组率为 90 .0 5 % ,扩增后的文库总滴度为 1.5 6× 10 7pfu μL。用PCR方法对总文库和随机挑取的噬菌斑进行扩增 ,表明λZiplox载体中已含有大小为 0 .5～ 8.0kb的cDNA插入片段 ,并在 2 .5kb、1.1kb和 0 .75kb处cDNA插入片段相对集中。所有分析结果表明 ,采用特定的发育时期和特定的组织提取的mRNA构建而成的cDNA表达文库具有较高频率的利它素基因片段和合适的滴度 ,定向插入技术并结合λZiplox噬菌体的表达功能 ,使构建的cDNA文库具有表达外源DNA的功能 ,有利于进行利它素基因的免疫筛选。

关 键 词：Telenomus theophilae host recognition kairomone cDNA library 

分 类 号：Q966[生物学—昆虫学] Q963