摘要
研究重组痘苗病毒对不同哺乳动物细胞的感染效率及表达水平,可为痘苗病毒表达系统宿主细胞的正确选择提供依据.本研究利用重组绿色荧光蛋白基因的痘苗病毒WR-EGFP同时感染不同的哺乳动物细胞株,利用流式细胞仪检测EGFP的表达强度.共使用20种哺乳动物细胞株,其中10种人类组织细胞,2种猴组织细胞,8种小鼠组织细胞.结果表明,重组痘苗病毒WR-EGFP对鼠细胞系BHK21和人细胞系A-549的感染效率和表达效率最佳;整体看,痘苗病毒对多数灵长类动物细胞的感染效率和表达效率优于鼠细胞;对贴壁细胞的感染效率和表达效率明显优于悬浮细胞;但没有特别的组织偏嗜性.
Vaccinia virus was widely used as gene expressing vector in mammalian cell lines and candidate recombinant vaccine vector,thus parallel comparation of the infection and expression efficiencies of recombinant vaccinia virus in different mammalian cell lines is essential for correctly choicing suitable expressing host cell lines. In this study, different MOI of recombinant vaccinia virus WR-EGFP were used to infect several different mammalian cell lines simultaneously,after 48 h p. o. i. ,and flow cytometer was used to detect the intensity of green fluorescence emitted by GFP. Twenty cell lines, 2 monkey cell lines and 8 rodent cell lines. Results showed kidney cell BHK21 and human cell A-549 with the highest infection different mammalian cell lines were infected, including 10 human that recombinant vaccinia virus WR-EGFP could infect hamster efficiency and expression efficiency. For most primate cell lines, WR-EGFP has higher infection efficiency and expression efficiency than rodent cell lines,and both efficiencies are higher in adherent culture cell lines than in suspend culture cell lines,but no obvious partiality was observed for cells coming from different organ origin. At 24 h p. o. i. ,different MOI of WR-EGFP infected cell and expressed GFP with markedly different intensity,but at 48 h p. o. i., the difference became negligible,which implies lower dosage of virus and longer culture can be used to express target proteins. Comparing with baculoviruses, vaccinia virus has higher infection and expression efficiency in mammalian cell lines with lower MOI,but may lysis most cell lines after 72 h p. o. i. , which seldom happen when infecting with baculoviruses.
出处
《厦门大学学报（自然科学版）》
CAS
CSCD
北大核心
2005年第6期866-869,共4页
Journal of Xiamen University：Natural Science
基金
福建省科技重大专项(2004YZ01)资助
