伪狂犬病病毒Ea株UL31基因克隆、序列分析及其在大肠杆菌中的表达

以伪狂犬病病毒Ea株基因组DNA为模板,通过PCR扩增含UL31基因的1000bp片段,扩增产物克隆于pMD18＿T中,双脱氧末端终止法序列测定。通过OM1GA2 0软件包分析发现,Ea株UL31基因编码271个氨基酸,蛋白质分子量为30 38kD,与Ka株UL31基因核苷酸与氨基酸序列同源性均在98%以上。将α＿疱疹病毒亚科9个不同成员UL31同源基因编码的氨基酸序列进行多重比对分析,发现4个保守的功能性结构域。将该片段插入原核表达载体pGEX＿KG中GST下游,构建的原核表达质粒pGEX＿UL31在大肠杆菌BL32(DE3)中获得了高效表达,SDS＿PAGE结果显示,表达的融合蛋白质分子量为56kD。

疱疹病毒UL31基因及其编码蛋白研究进展

疱疹病毒是一类有囊膜的双链DNA病毒,基因组由长节段(L)和短节段(S)组成。UL31基因是基因组独特UL序列中的一个核心基因,具有典型的开放阅读框架,编码一种磷酸化蛋白。该蛋白与UL34蛋白相互作用,定位于细胞核边缘,与病毒核衣壳初期包膜形成有关,对病毒DNA的合成、包装和释放也有一定的作用。本文就UL31基因及编码蛋白的结构特点和基本功能等研究进展进行总结,以期为疱疹病毒UL31基因的深入研究提供指导。

Pseudo Rabies Virus Protein UL34 Interacted with UL31 to Disrupt the Human Lamin A

Pseudo rabies virus（PRV） egresses from the nucleus by budding from the inner nuclear membrane（INM）. The nuclear lamina forms a rigid meshwork of intermediate filaments underlying the INM. It remains unknown whether PRV infection induces the disruption of lamina. In this paper, it can be observed that nuclear Lamin A became fractured during PRV infection. UL34 was localized at the nuclear rim, but UL31 was accumulated in the nucleus as distinct patches. Interestingly, a part of UL31 was localized at the INM in the presence of UL34. Immunoprecipitation（IP） assay confirmed that PRV UL31 and UL34 interacted in the transfected cells. Importantly, the co-expression of UL31 and UL34 directly disrupted Lamin A, resembling that observed during PRV infection. In conclusion, PRV infection induces the disruption of Lamin A, and UL34 and UL31 play a critical role in the disruption of Lamin A.