CHEN M ing—ruing, WU Dong, YUAN Li, CHEN Xin—wen and HU Zhi—hong. Prokaryotic Expression of Three bro Genes from Helicoverpa armigera Singlenucleocapsid Nucleopolyhedrovirus and Generation of Antibodies[J]. Virologica Sinica, 2003, 18(6): 597-602.
Citation:
CHEN M ing—ruing, WU Dong, YUAN Li, CHEN Xin—wen, HU Zhi—hong.
Prokaryotic Expression of Three bro Genes from Helicoverpa armigera Singlenucleocapsid Nucleopolyhedrovirus and Generation of Antibodies .VIROLOGICA SINICA, 2003, 18(6)
: 597-602.
棉铃虫单核衣壳核多角体病毒3个bro基因的原核表达与抗体制备
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摘要
摘要：根据棉铃虫单核衣壳核多角体病毒(HaSNPV)基因组全序列，设计引物，用PCR的方法扩增得到bro-a、bro-b 和bro．C三个全长基因。将这三个基因的片段分别克隆至原核表达载体pProExHTb，经IFIX3诱导，在E．coliDH50 菌株中得到了目的基因的高效表达。表达的目的蛋白大小分别为32kDa、64kDa和58kDa，经SDS-PAGE分离纯 化，免疫新西兰大白兔制备了多克隆抗体。抗体经1：2500倍稀释后用于WestemBlot分析，获得特异性显色信 号，所制备的三种抗体适合用作bro基因的功能的进一步研究。
Abstract：Three baculovirus repeated open reading frame(bro)genes were amplified by PCR from thegenome DNA of HaSNPV．The PCR products were cloned into pBluescript KS(+)plasmid．The geneswere introd uced into the expression vector pProExHTb．After IFFG induction．the E coli DH5a straincontaining each of the recombinant plasmids expressed proteins with molecular weights of 32 kDa、64kDa an d 58 kDa,respectively,which were in agreement with the prospeetation．Th e purifiedrecombinant proteins were used to immune the rabbits separately．Western blot an alysis using themulticlonal an tibodies derived from the rabbits indicated that these an tibodies could react specificallywith thetargetproteinsan dwere suitableto beusedforfurtherfunctional an alysisofthebrogenes．
Prokaryotic Expression of Three bro Genes from Helicoverpa armigera Singlenucleocapsid Nucleopolyhedrovirus and Generation of Antibodies
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1.
Joint-lab ofInvertebrate Virology and Key Laboratory of Molecular Virology,Wuhan Institute of Virology,Chinese AcademyofSciences,Wuhan430071，China
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Abstract
Abstract：Three baculovirus repeated open reading frame(bro)genes were amplified by PCR from the genome DNA of HaSNPV．The PCR products were cloned into pBluescript KS(+)plasmid．The genes were introd uced into the expression vector pProExHTb．After IFFG induction．the E coli DH5a strain containing each of the recombinant plasmids expressed proteins with molecular weights of 32 kDa、64 kDa an d 58 kDa,respectively,which were in agreement with the prospeetation．Th e purified recombinant proteins were used to immune the rabbits separately．Western blot an alysis using the multiclonal an tibodies derived from the rabbits indicated that these an tibodies could react specifically with thetargetproteinsan dwere suitableto beusedforfurtherfunctional an alysisofthebrogenes．
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