Construction of Agrotis segetum granulosis virus cDNA Library

ZHANG Wei; WANG Wei; AI Xiu-lian”; XIE Yu-qing; FEN Huai-rong

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February 2003

ZHANG Wei, WANG Wei, AI Xiu-lian”, XIE Yu-qing and FEN Huai-rong. Construction of Agrotis segetum granulosis virus cDNA Library[J]. Virologica Sinica, 2003, 18(1): 27-30.

Citation: ZHANG Wei, WANG Wei, AI Xiu-lian”, XIE Yu-qing, FEN Huai-rong. Construction of Agrotis segetum granulosis virus cDNA Library .VIROLOGICA SINICA, 2003, 18(1) : 27-30.

黄地老虎颗粒体病毒cDNA文库的构建

1.
f新疆农科院微生物应用研究所基因工程实验室，新疆乌鲁木齐830000

摘要

：以感染黄地老虎颗粒体病毒 Agrotis segetum ganulosis virus，AsGV)的黄地老虎幼虫为材料提取总RNA、分离mRNA，并反转录合成cDNA，构建了包括黄地老虎(Agrotis segetum，As)幼虫和黄地老虎颗粒体病毒的 cDNA文库。用EcoR I和HindIII限制性内切酶酶切AsGV基因组DNA，制备地高辛探针．与上述总RNA、mR． NA、cDNA杂交，从文库中筛选出AsGV的阳性克隆1081个，经cDNA测序，cDNA编码序列与基因组编码序列相符。并根据基因组阅读框序列合成引物，PCR扩增出59个阅读框的编码基因，也完全与基因组序列相符。
- 黄地老虎颗粒体病毒
- , 基因组序列
- , cDNA文库
- , 阳性克隆

Construction of Agrotis segetum granulosis virus cDNA Library

1.
Detector ofGene Biotechnology Laboratory，Institute ofMicrobiology xiaj~ gAcademy ofAgricultural Science， Xinjiang Wulumuqi 830000，China

Abstract

Abstract：A general cDNA library of Agrotis segetum granulosis virus(ASGV)which was from infected A grotis segetum(As)l arva has been constructed．The first and second cDNA strands were synthesized by AMV reverse transcriptase from mRNA which was isolated from total RNA of infected As larva． So the general cDNA library includes AsGV’S and AS’S， Th en the library of AsGV included 108 1 positive clones were screened by spot hybridization， Th e labeled AsGV genome DNA probe With DIG kit was cut by restriction enzyme of EcoR I and Hind 11． Th e sequence of positnive clones accord with the genome of AsGV，and there are 59 coding sequence which were amplified with synthetic primer of As— GV genome according entirely with the genome of AsGV．
- Agrotis segetum granulos virus
- , Genome sequence
- , cDNA library
- , Positive clone

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通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142

Construction of Agrotis segetum granulosis virus cDNA Library

1. Detector ofGene Biotechnology Laboratory，Institute ofMicrobiology xiaj~ gAcademy ofAgricultural Science， Xinjiang Wulumuqi 830000，China

Abstract: Abstract：A general cDNA library of Agrotis segetum granulosis virus(ASGV)which was from infected A grotis segetum(As)l arva has been constructed．The first and second cDNA strands were synthesized by AMV reverse transcriptase from mRNA which was isolated from total RNA of infected As larva． So the general cDNA library includes AsGV’S and AS’S， Th en the library of AsGV included 108 1 positive clones were screened by spot hybridization， Th e labeled AsGV genome DNA probe With DIG kit was cut by restriction enzyme of EcoR I and Hind 11． Th e sequence of positnive clones accord with the genome of AsGV，and there are 59 coding sequence which were amplified with synthetic primer of As— GV genome according entirely with the genome of AsGV．