华南农业大学学报2011，Vol.32Issue(4)：96-100,5.
表面展示猪圆环病毒2型衣壳蛋白的重组杆状病毒的构建
Construction of Recombinant Baculovirus Surface-Displayed the Capsid Protein of Porcine Circovirus Type 2
摘要
Abstract
The open capsid(Cap)protein gene without nuclear localization signal (NLS) of porcine circo-virus 2(PCV2)was amplified by PCR and sub-cloned into pBACsurf-1 between the upstream gp64 signal sequence and downstresm gp64 mature domain. The fusion gene containing dCap and gp64 was inserted into pcDNA3.1 ( + ) to construct the recombinant plasmid pcDNA3. L-gp64-dCap. A fragment of the CMV promoter-gp64-dCap fusion gene cassette was excised from pcDNA3. L-gp64-dCap and inserted into the baculovirus transfer vector pFastBac-V to construct the recombinant plasmid pFastBac-dCap-V. It was followed by the transformation of plasmid into Escherichia coli DHlOBac competent cells to obtain the recombinant shuttle vector Bacmid-dCap-V. Eventually, the recombinant Bacmid was transfected into Sf9 cells to produce the recombinant baculovirus Ac-dCap-V using the LipofectamineTM 2000. The Sf9 cells produced cytopathic after infection of recombinant baculoviruses Ac-dCap-V. Indirect immunofluoresent assay demonstrated that the Ac-dCap-V efficiently transducted the mammalian cells in vitro and the dCap protein was expressed successfully.关键词
杆状病毒表面展示系统/猪圆环病毒2型/dCap基因Key words
baculovirus surface display/ porcine circovirus 2/ dCap gene分类
农业科技引用本文复制引用
程晓亮,林文耀,叶煜,陈筱薇,严常燕,廖明,樊惠英..表面展示猪圆环病毒2型衣壳蛋白的重组杆状病毒的构建[J].华南农业大学学报,2011,32(4):96-100,5.基金项目
863计划(2006AA10A205) （2006AA10A205）
教育部新世纪优秀人才支持计划(NCET-06-0752) （NCET-06-0752）
国家自然科学青年基金(30800826) （30800826）
广东省博士启动基金(8451064201001131) （8451064201001131）
农业微生物学重点实验室开发课题(20090010) （20090010）
华南农业大学校长基金(5500-K08240) （5500-K08240）
