中华卵索线虫cDNA文库的构建  

The cDNA library construction of Ovomermis sinensis

作　　者：陈柳惠[1] 陈冲[1] 焦振龙[1] 王国秀[1] 

机构地区：[1]华中师范大学生命科学学院遗传与整合生物学湖北省重点实验室,武汉430079

出　　处：《华中师范大学学报（自然科学版）》2014年第3期404-407,共4页Journal of Central China Normal University：Natural Sciences

基　　金：国家自然科学基金项目(30870350)

摘　　要：中华卵索线虫（Ovomermis sinensis）是一种宝贵的昆虫天敌,宿主广泛,其寄生率即等于宿主的死亡率,生防潜力极大.鉴于cDNA文库在保存稀有物种基因资源与目的基因筛选及其功能研究等方面的优势,实验以中华卵索线虫为材料,采用SMART技术构建其cDNA文库.实验结果表明：滴度为1.16×10^9 cfu·mL^-1,包含6.0×10 ^6个克隆.从原始文库中随机挑16个单菌落过夜培养,提取质粒,经XhoⅠ和PstⅠ双酶切,琼脂糖凝胶电泳结果表明插入片段大小主要集中在0.5～2.0 kb之间,文库重组率接近100％.文库的各项指标均达要求,可用于全长cDNA的筛选,为新基因的获得及其结构功能研究提供了可靠材料,也为今后的生物防治等研究奠定了基础.Ovomermis sinensis is insect parasitic nematodes,its host is very widely,and the parasitism is equal with the insect mortality.So it＇s a valuable insect natural enemies.In order to save the gene resources of the O.sinensis and study the function of genes,we constructed the cDNA library of O.sinensis by SMART（switching mechanism at 5＇end of RNA transcript）technique.The results show that：The titer of the cDNA library was 1.16 × 10^9 Cfu · mL^-1,the capacity of the library was 6.0 × 10^6 clones.Picked 16 clones randomly,extracted their plasmids and identified by the restriction enzyme of Xho Ⅰ and Pst Ⅰ.The gel electrophoresis result indicated that the fragments ranged from 0.5～2.0 kb and the recombination rate was nearly 100%.These results showed that the cDNA library was constructed successfully and it could be used to screen the new genes and to research the application of O.sinensis in the biological control.

关 键 词：中华卵索线虫 CDNA文库 SMART技术 

分 类 号：Q812[生物学—生物工程]