棉铃虫核型多角体病毒泰国株ORF27全长基因序列分析及其截短型基因的原核表达  

Analysis of ORF27 full length sequence of Thai isolated Helicoverpa armigera nucleopolyhedrovirus and prokaryotic expression of truncated ORF27 gene

作　　者：寻萌[1] POOMPUTSA Kanokwan 

机构地区：[1]西安交通大学医学院免疫与病原生物学系 [2]泰国玛古国王理工大学动物细胞实验室

出　　处：《西安交通大学学报（医学版）》2010年第3期293-298,共6页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：联合国"生物工学校际研究生课程"项目~~

摘　　要：目的研究棉铃虫核型多角体病毒泰国株ORF27蛋白的功能,阐释其在传代效应中可能发挥的分子机制。方法用PCR方法从棉铃虫核型多角体病毒泰国株基因组中扩增ORF27基因;使用ProtParam、TMpred、SignalP、ScanProsite等生物信息学软件分析其理化特性、亚细胞定位、信号肽、跨膜结构、功能位点等;并将其氨基端截短型基因进行原核表达。结果ORF27基因序列全长768 nt,编码255 aa,分子质量约29.5 ku。生物信息学软件预测其具有多个功能基序。截短型重组载体表达出分子质量约35 ku的外源蛋白,主要以包涵体形式存在。结论棉铃虫核型多角体病毒ORF27蛋白可能具有非常广泛的生物学活性,调控多种细胞信号传导通路,调节细胞凋亡和细胞周期,参与传代效应的发生。Objective To understand the functions of ORF27 protein of the Thai isolated Helicoverpa armigera nucleopolyhedrovirus（HaNPV-Th） and explain the possible molecular mechanism of HaNPV-Th in passage effect.Methods ORF27 gene was cloned and amplified by polymerase chain reaction（PCR） from HaNPV-Th genome.The physical-chemical properties,subcellular localization,signal peptide,transmembrane structure and functional motifs were analyzed by the software of ProtParam,TMpred,SignalP and ScanProsite.The truncated ORF27 gene which lacked parts of the amino-terminal was expressed by bacterial expression system.Results The full length sequence of ORF27 was 768bp long and encoded 255aa with a molecular weight of 29.5ku.Several functional motifs on ORF27 were predicted by bioinformatics software.The truncated ORF27 recombinant expressed 35ku target protein mainly in the form of inclusion body.Conclusion ORF27 protein of HaNPV-Th is presumed to possess extensive biological activities.It may regulate various cellular signaling pathways,apoptosis and cell cycle,which are involved in passage effect.

关 键 词：棉铃虫核型多角体病毒泰国株(HaNPV-Th) 传代效应 ORF27 序列分析 原核表达 

分 类 号：S181[农业科学—农业基础科学] S188