摘要
从构建的重组质粒pLEX-C中高保真PCR获得编码登革2型病毒43株C基因(D2C)的DNA片段,通过基因重组的方法将其克隆入真核表达载体pcDNA6/V5-His获得了重组真核表达载体pc/D2C。经电穿孔的方法转染BHK21细胞后,分别通过RT-PCR、免疫荧光和Western印迹鉴定表达的蛋白。结果重组蛋白在BHK21细胞中获得表达,表达的蛋白主要存在于胞浆中,并具有较好的抗原性,能够被抗登革病毒衣壳蛋白单克隆抗体特异识别。此研究为深入了解登革病毒衣壳蛋白在病毒复制及组装过程中的生物学功能奠定了基础。
The D2C gene fragments were amplified by high fidelity PCR from the constructed plasmid pLEX-C and then cloned into enkaryotic expression vector pcDNA6/V5-His to generate the desired recombinant plasmid pc/D2C. After transfected into the mammalian BHK21 cells by electroporation, the expression of recombinant proteins were identified by RT-PCR, indirect immunofluorescence and Western blot. The results of IFA showed the existence of D2C in the cytoplasm. The results of Western blot indicated the expressed D2C could react with the Mab 8H8 that specifically identified the capsid protein of dengue-2 virus. This work laid the foundation of interests on the biological function of capsid protein during the life cycle of dengue virus.
出处
《生物技术通讯》
CAS
2005年第6期595-597,共3页
Letters in Biotechnology
基金
国家自然科学基金项目(30100006
30570084)
关键词
登革病毒
衣壳蛋白
真核表达
dengue virus
capsid protein
enkaryotic expression
