诱导表达极早期基因ie-1反义序列抑制杆状病毒复制的试验  

A Preliminary Study on Inhibiting the Replication of Baculovirus by Induced Expression of Immediate-early Gene ie-1 Antisense Sequence

作　　者：刘子瑜[1] 车凤玉[1] 张业顺[2] 夏定国[2] 张国政[2] 韦亚东[2] 

机构地区：[1]江苏科技大学生物与化学工程学院,江苏镇江212018 [2]中国农业科学院蚕业研究所,江苏镇江212018

出　　处：《蚕业科学》2012年第5期850-855,共6页ACTA SERICOLOGICA SINICA

基　　金：国家自然科学基金项目(No.30871829)

摘　　要：在昆虫杆状病毒的级联复制中,极早期基因ie-1所编码的IE-1蛋白具有重要功能,其中极晚期多角体蛋白基因启动子polh受其调控激活。通过建立具有启动子polh及ie-1长194 bp的反义序列(ie-1-r)的转录载体,使杆状病毒在宿主细胞进行自身复制时受到抑制。试验结果表明,当转入重组转录载体polh/ie-1-r的宿主细胞受携带荧光素酶(luciferase)基因的杆状病毒感染时,能够被诱导激活细胞内多角体蛋白基因启动子转录其携带的ie-1的反义序列RNA,从而部分抑制病毒本身的复制,宿主细胞在病毒感染第4～5天,其荧光素含量与对照RLU(relative light unit)相比下降约1×108个单位,抑制病毒的效果在时间上呈现一定的滞后性。试验结果提示:构建类似的病毒诱导表达系统,通过早期基因激活晚期基因,晚期基因转录抑制早期基因表达的策略,可以对病毒复制进行抑制。During the replication of insect baculovirus, IE-1 protein, coded by immediate early gene ie-1, has impor- tant roles such as to regulate the activation of late polyhedrin gene promoter polh. in present study, we constructed a transcription vector carrying the promoter polh and a 194 bp antisense sequence of ie-1 （termed as ie-l-r） to restrain the self-replication of baculovirus in host cells. The experimental results showed that, when transformed host cells har- boring the recombinant transcription vector polh/ie-l-r were infected by baculovirus carrying luciferase gene, promoter polh of polyhedrin gene could be induced and activated to transcribe the RNA of ie-1 antisense sequence, resulting in a partial inhibition to self-replication of baculovirus. Compared to relative light unit （RLU） of the control, the fluorescein content in host cells declined about 1 x 108 units on day 4 to day 5 after virus infection, showing certain time-lag effecton inhibition to virus replication. These experimental results indicate that virus replication could be inhibited through constructing similar virus-induced expression system based on the strategy of activating late gene u- sing early gene and inhibiting expression of early gene through transcription of the bate gene.

关 键 词：多角体蛋白基因启动子 ie-1基因 反义序列 病毒复制 抑制 

分 类 号：S884.5[农业科学—特种经济动物饲养] Q78[农业科学—畜牧兽医]