【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。更多还原

【Abstract】 During the replication of insect baculovirus,IE-1 protein,coded by immediate early gene ie-1,has important 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-1-r) to restrain the self-replication of baculovirus in host cells.The experimental results showed that,when transformed host cells harboring the recombinant transcription vector polh/ie-1-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 1×108 units on day 4 to day 5 after virus infection,showing certain time-lag effect on 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 using early gene and inhibiting expression of early gene through transcription of the late gene.更多还原