【摘要】 目的构建携带人乳头瘤病毒(HPV)结构蛋白L1和白色念珠菌甘露糖蛋白(CaMp65)细胞毒性T细胞表位(CTL)基因的重组质粒,并在杆状病毒-昆虫细胞系统中进行表达。方法分别设计包含HPV6bL1和CaMp65CTL145-153的引物,经PCR扩增嵌合基因HPV6bL1/CaMp65CTL145-153,并进一步将其插入杆状病毒表达载体pFastBacTM1。在E.coliDH10Bac中组装成重组杆状病毒,在昆虫细胞sf-9中表达嵌合蛋白,并经SDS-PAGE和Westernblot分析鉴定。结果HPV6bL1/CaMp65CTL145-153嵌合蛋白在昆虫细胞sf-9中得到了表达,表达产物的相对分子质量为56000,与HPV6bL1单抗能产生特异性反应。结论HPV6bL1/CaMp65CTL145-153嵌合蛋白在杆状病毒-昆虫细胞系统中得到了成功表达,为防治HPV和白色念珠菌感染的基因工程疫苗的研究奠定了基础。更多还原

【Abstract】 Objective To construct recombinant plasmid carrying the genes of human papillomavirus(HPV)type 6b structural protein L1 and CTL epitope of Candida albicans(Ca)mannoprotein(Mp)and express in baculovirus-insect cell system.Methods Chimeric gene HPV6b L1/CaMp65 CTL145-153 was amplified by PCR with designed primers and inserted into baculovirus transfer vector pFastBacTM1,then transformed to E.coli DH10Bac.The constructed recombinant baculovirus rBacmid/HPV6b L1/CaMp65 CTL145-153 was tranfected to insect cell strain sf-9,and the expressed product was identified by SDS-PAGE and Western blot.Results HPV6b L1/CaMp65 CTL145-153 chimeric protein with a relative molecular mass of 56 000 was expressed in sf-9 cells and showed specific reaction with McAb against HPV6b L1.Conclusion HPV6b L1/CaMp65 CTL145-153 chimeric protein was successfully expressed in baculovirus-insect cell system,which laid a foundation of developing recombinant vaccine for prevention and treatment of HPV and Candida albicans infections.更多还原