Incorporation of GP64 into Helicoverpa armigera nucleopolyhedrovirus enhances virus infectivity in vivo and in vitro
Incorporation of GP64 into Helicoverpa armigera nucleopolyhedrovirus enhances virus infectivity in vivo and in vitro
复制标题
将 GP64 掺入棉铃虫核多角体病毒可增强病毒体内和体外的感染性。
DOI：
10.1099/vir.0.046458-0
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发表时间：
2012-12-01
影响因子：
3.8
通讯作者：
Deng, Fei
中科院分区：
文献类型：
--
作者：
Shen, Shu;Gan, Yinyin;Deng, Fei
杆状病毒的包膜融合蛋白，来自I组核型多角体病毒（NPV）的糖蛋白GP 64或来自II组NPV和颗粒病毒的F蛋白，对于杆状病毒的形态发生和感染性是必需的。F蛋白被认为是祖先杆状病毒包膜融合蛋白，而GP 64是杆状病毒中的更新的进化引入，并且表现出比F蛋白更高的融合活性。每种融合蛋白都是相应病毒在幼虫组织内传播感染所必需的。构建了表达苜蓿银纹夜蛾核型多角体病毒GP 64的重组棉铃虫核型多角体病毒（HearNPV）vHaBac-gp 64-egfp，并测定了其体内外感染性。通过一步生长曲线测定病毒滴度和通过定量PCR测定病毒DNA拷贝数的分析表明，vHaBac-gp 64-egfp比对照vHaBac-egfp具有更强的体外感染性。将多角体蛋白基因（polh）重新引入重组病毒中，生物测定表明，与vHaBac-egfp-polh对照相比，vHaBac-gp 64-polh加速了感染幼虫的死亡率，并且vHaBac-gp 64-polh的LC（50）（半数致死浓度）降低到vHaBac-egfp-polh的约20%。 因此，GP 64掺入HearNPV出芽病毒粒子改善了病毒在体内和体外的感染性。该双价病毒的构建将进一步提高杆状病毒在基因治疗和生物防治等领域的应用。
The envelope fusion proteins of baculoviruses, glycoprotein GP64 from group I nucleopolyhedrovirus (NPV) or the F protein from group II NPV and granulovirus, are essential for baculovirus morphogenesis and infectivity. The F protein is considered the ancestral baculovirus envelope fusion protein, while GP64 is a more recent evolutionary introduction into baculoviruses and exhibits higher fusogenic activity than the F protein. Each of the fusion proteins is required by the respective virus to spread infection within larval tissues. A recombinant Helicoverpa armigera NPV (HearNPV) expressing GP64 from Autographa californica multiple nucleopolyhedrovirus, vHaBac-gp64-egfp, was constructed, which still retained the native F protein, and its infectivity was assayed in vivo and in vitro. Analyses by one-step growth curve to determine viral titre and by quantitative PCR to determine viral DNA copy number showed that vHaBac-gp64-egfp was more infectious in vitro than the control, vHaBac-egfp. The polyhedrin gene (polh) was reintroduced into the recombinant viruses and bioassays showed that vHaBac-gp64-polh accelerated the mortality of infected larvae compared with the vHaBac-egfp-polh control, and the LC(50) (median lethal concentration) of vHaBac-gp64-polh was reduced to approximately 20 % of that of vHaBac-egfp-polh. Therefore, incorporation of GP64 into HearNPV budded virions improved virus infectivity both in vivo and in vitro. The construction of this bivalent virus with a more efficient fusion protein could improve the use of baculoviruses in different areas such as gene therapy and biocontrol.