蓖麻蚕核型多角体病毒基因gp64的克隆及荧光定量表达分析  

Cloning and expressional analysis by fluorescent quantitative PCR of Philosamia cynthia ricini nuclear polyhedrosis virus gp64 gene

作　　者：钱荷英[1,2] 徐安英[1,2] 张月华[1,2] 孙平江[1,2] 高坤[1] 郭锡杰[1,2] 

机构地区：[1]江苏科技大学,江苏镇江212003 [2]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《河北农业大学学报》2013年第6期76-82,共7页Journal of Hebei Agricultural University

基　　金：国家基础研究计划(2012CB114600)

摘　　要：Gp64是杆状病毒CRV(Cell-released virus)的主要嚢膜蛋白,在CRV借吸附和内吞作用侵入细胞过程中起关键作用,能促进CRV嚢膜与胞饮体膜之间的融合从而促进杆状病毒对宿主细胞的吸附。本试验克隆了蓖麻蚕核型多角体病毒(Philosamia cynthia ricini nuclear polyhedrosis virus,PcrNPV)的gp64基因,序列分析表明该基因ORF全长1 530bp,编码509个氨基酸残基,其相应的GP64蛋白质分子量为58.46kD,等电点为5.59;同源性分析表明,PcrNPV的GP64蛋白与柞蚕核型多角体病毒(Antheraea pernyi nuclear polyhedrosis virus,ApNPV)的GP64同源性高达99%,而与家蚕核型多角体病毒(Bombyx mori nuclear polyhydrosis virus,BmNPV)的GP64仅为76%;荧光定量分析表明,PcrNPV的gp64基因在不同蓖麻蚕品种的中肠、血液及其脂肪体中均有相同的增殖趋势,但在不同品种的各组织中的增殖量不同,增殖量多的品种与感染试验中相对易感的品种一致。GP64 is the major envelope membrane protein of the cell released virus （CRV）, and play a key role in penetrating a cell of CRV during endocytosis and adsorption. The gp64 in PcrNPV was cloned, and its ORF contains 1 530 bp and encodes a protein of 509 amino acid residues with a calculated molecular weight of 58.46 kD and isoelectric point of 5.59. The ho- mology analysis of the gp64 gene sequences indicated that the deduced amino acid sequence of PcrNPV shared homology of 99% and 76 % with ApNPV and BmNPV, respectively. Real-time quantitative RT-PCR results indicated that the trends of expression of GP64 in midgut, blood and fat body of different varieties were similar, but the relative amount of expression in differ-ent tissues was different. The variety which showed higher expression was consistent with the more susceptive variety in the experiments.

关 键 词：蓖麻蚕核型多角体病毒 GP64 基因克隆 荧光定量PCR 

分 类 号：S685.26[农业科学—观赏园艺]