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葱蝇冬滞育蛹的全长cDNA文库的构建
引用本文:黎万顺,冯国忠,陈斌,何正波.葱蝇冬滞育蛹的全长cDNA文库的构建[J].昆虫知识,2010,47(1):53-58.
作者姓名:黎万顺  冯国忠  陈斌  何正波
作者单位:昆虫与分子生物学研究所重庆高校生物活性物质工程研究中心,重庆高校动物生物学重点实验,室重庆,400047
基金项目:国家自然科学基金(No.30870340);;重庆市自然科学基金(No.CSTC2008BB1365)资助;;重庆市自然科学基金重点项目(No.CSTC2008BA5030)
摘    要:葱蝇Delia antiqua(Meigen)具有冬滞育的特性且与黑腹果蝇Drosophila melanogaster近缘。该研究目的在于构建葱蝇冬滞育蛹的全长cDNA文库,为进一步的冬滞育专化基因筛选、克隆、和表达分析奠定基础。利用RNAiso试剂盒提取葱蝇冬滞育蛹的总RNA,采用SMART技术合成全长cDNA,经SfiⅠ酶切消化后,将cDNA克隆到质粒载体pDNR-LIB。经鉴定,原始文库的滴度为2.4×107cfu/mL。随机挑取15个克隆,经PCR快速鉴定,插入片段大小在0.5~3kb之间,平均大小在1kb左右,重组率达100%。结果表明该研究获得高质量的葱蝇冬滞育蛹cDNA文库。

关 键 词:葱蝇  冬滞育蛹  全长cDNA文库  SMART技术

Construction of full-length cDNA library of winter-diapause pupae of the onion maggot, Delia antique.
LI Wan-Shun,FENG Guo-Zhong,CHEN Bin,HE Zheng-Bo.Construction of full-length cDNA library of winter-diapause pupae of the onion maggot, Delia antique.[J].Entomological Knowledge,2010,47(1):53-58.
Authors:LI Wan-Shun  FENG Guo-Zhong  CHEN Bin  HE Zheng-Bo
Institution:Institute of Entomology and Molecular Biology/a>;Chongqing Engineering Research Center of Bioactive Substances/a>;Chongqing Key Laboratory of Animal Biology/a>;Chongqing Normal University/a>;Chongqing 400047/a>;China
Abstract:The onion maggot,Delia antiqua(Meigen),is characterized by winter-diapause and is close to Drosophila melanogaster in phelogenetics. This study aims to construct full-length cDNA library of winter-diapause pupae of D. antique,in order to provide a base for further screening,cloning and expression analysis of winter-diapause-specific genes. In this study,total RNA was extracted from winter-diapause pupae of D. antiqua using RNAiso kit. Double-stranded cDNAs were synthesized with SMART technique and digested ...
Keywords:Delia antiqua  winter-diapause pupae  full-length cDNA library  SMART technique  
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