摘要
对含伪狂犬病病毒 Ea株 g C全基因的质粒 p UC1 .75进行亚克隆 ,将其完整编码区置于真核表达载体 pc DNA3.1 +的 HCMV启动子 /增强子下游 ,构建了 g C基因真核表达质粒 pc DNA-g C.脂质体转染 IBRS- 2细胞 ,在 G41 8抗性选择下 ,获得多个阳性克隆细胞系 .经 ELISA筛选反应最强的阳性克隆细胞系 ,进一步用间接免疫荧光检测证实 g C基因在 IBRS- 2细胞中得到了正确表达并分布在细胞膜 .以 1 0 0个蚀斑形成单位的伪狂犬病病毒分别接种表达 g C的细胞系和空白载体转染细胞系 .通过测定蚀斑数发现 ,表达 g C的细胞系对病毒的感染具有抑制作用 ,平均抑制率达 59.4%± 1 .3% .
A eukaryotic expression plasmid pcDNA\| gC containing the full\|length glycoprotein C (gC) gene of pseudorabies virus Ea strain was constructed.pcDNA\| gC was transfected into IBRS\|2 cells and several clone cells were obtained in the selection of G418.The best clone was screened via enzyme\|linked immunosorbent assay (ELISA).Expressed protein was further detected by indirect immunofluorescence assay.When approximately 100 plauqe\|forming units(pfu) of PrV were inoculated the cell line expressing gC and the control cell respectively,the numbers of plaques showed that the cell line expressing gC could inhibited virus infection and the inhibition rate was up to 59 4%±1 3%.
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2001年第2期198-202,共5页
Chinese Journal of Biochemistry and Molecular Biology
基金
国家自然科学基金!资助项目 (No.396 70 5 5 5
39970 5 5 9)&&
