博尔纳病毒P24转染后不同时期核酸和蛋白的表达及细胞定位  

Expression and cellular localization of Borna disease virus P24 in different periods after transfection

作　　者：徐鸣明[1] 张英英[1] 展群岭[1] 何丰[1] 张亮[1] 金戈[1] 宋哲[1] 李亚军[2] 李雷雷[3] 谢鹏[1,3] 

机构地区：[1]重庆医科大学附属第一医院神经内科,重庆400016 [2]西安医学院附属医院神经内科,西安710077 [3]重庆医科大学神经科学中心,重庆400016

出　　处：《第三军医大学学报》2010年第6期533-536,共4页Journal of Third Military Medical University

基　　金：国家高技术研究发展计划(863计划,2006AA02Z196)~~

摘　　要：目的通过观察博尔纳病毒(borna disease virus,BDV)磷蛋白(P24)及其编码核酸不同时期的表达和细胞定位,探索BDV潜伏感染的机制。方法构建包含BDV GFP-P24质粒的OL细胞模型,检测并比较不同时期P24核酸和蛋白的表达;对构建的OL细胞模型和各种阴性对照进行逆转录原位PCR的检测,观察P24蛋白编码核酸的细胞定位;使用倒置荧光显微镜观察OL细胞中P24蛋白的细胞定位。结果成功构建了含BDV GFP-P24质粒的OL细胞,发现转染后20代以内细胞P24核酸和蛋白的表达差异不显著(P>0.05);BDV的P24重组蛋白基因始终定位于细胞核,其蛋白早期在细胞核出现,反复传代约15次后可同时在细胞核和细胞质内出现。结论BDV的重组蛋白P24在病毒感染过程中起关键作用,其编码基因始终存在于宿主细胞核内,但表达的蛋白可以在一定时期通过核膜进入细胞质。Objective To identify the cellular localization of Borna disease virus （BDV） P24 by in situ RT-PCR in different periods after transfection and explore the mechanism of BDV latent infection. Methods Oligodendrocyte （OL cells） were transfected by plasmid BDV GFP-P24, and then tested by RT- PCR and ELISA in different periods after transfection in order to compare different expressions of nucleic acid and protein. In situ RT-PCR and fluorescence microscopy were used to identity cellular localization of BDV P24 nucleic acid and protein in different periods. Results BDV P24 transfected OL model was successfully constructed. The expression of BDV P24 nucleic acid and protein showed there was no statistics difference （ P 〉 0. 05 ） in different periods in 20 passages. The BDV P24 gene was located in the cellular nucleus but BDV P24 protein in the cellular nucleus first and then in both nucleus and cytoplasm after 15 passages. Conclusion BDV P24 gene functions in the cellular nucleus and P24 protein can transport to the cytoplasm of OL cells at 15 passages or so after transfection.

关 键 词：博尔纳病 逆转录原位PCR 磷蛋白 感染 

分 类 号：R373.9[医药卫生—病原生物学] R394-33[医药卫生—基础医学]