|
|
|
新城疫病毒F48E9株M NP F和HN基因的真核表达
|
|
|
|
闻晓波，闫丽辉，曹殿军，刘培欣，刘春国，步志高
|
|
|
|
(中国农业科学院 哈尔滨兽医研究所 兽医生物技术国家重点实验室,黑龙江 哈尔滨150001)
|
|
|
|
摘要：
|
|
将新城疫病毒（NDV）F48E9株的M、NP、F和HN基因克隆到真核细胞表达载体pCAGG上，经酶切、PCR鉴定和序列分析，分别筛选出了含有M、NP、F和HN基因的重组质粒，命名为pCAGG-M、 pCAGG-NP、pCAGG-F和pCAGG-HN。纯化后的重组质粒通过脂质体单独转染HeLa细胞，经间接免疫荧光试验检测到了M、NP、F和HN蛋白的表达。pCAGG-M、 pCAGG-NP、pCAGG-F和pCAGG-HN重组质粒组合共转染HeLa细胞48h后，可以观察到明显的细胞融合现象。试验结果表明，NDV F48E9株的M、NP、F和HN蛋白均在HeLa细胞内得到了成功表达，同时证明在该表达系统中F蛋白不足以诱导细胞融合。 |
|
关键词：
|
|
新城疫病毒；M基因；NP基因；F基因；HN基因；真核细胞表达；病毒样颗粒 |
|
中图分类号：S 852.659.5:Q 786 文献标识码：
A 文章编号：1673-4696(2006)06-0429-05 |
|
|
Eukaryotic expression of M, NP, F and HN genes of NDV strain F48E9 |
|
|
|
WEN Xiao-bo, YAN Li-hui, CAO Dian-jun, LIU Pei-xin, LIU Chun-guo, BU Zhi-gao |
|
|
|
(National Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute,
Chinese Academy of Agricultural Sciences, Harbin 150001, China )
|
|
|
Abstract: M, NP, F and HN genes amplified from Newcastle disease virus F48E9 strain were cloned into the eukaryotic expression vector pCAGGS, respectively. The positive recombinant plasmids were named pCAGG-M, pCAGG-NP, pCAGG-F and pCAGG-HN after identification by restriction digestion, PCR amplification and sequencing analyses. Then the recombinant plasmids were amplified, purified and transfected into HeLa cells. Expression of M, NP, F and HN genes were detected and confirmed by the indirect immunofluorescence analysis. The syncytium formation was observed in the HeLa cells co-transfected with pCAGG-M, pCAGG-NP, pCAGG-F and pCAGG-HN recombinant plasmids at 48h post transfection. These results indicated that all four recombinant proteins were successfully expressed in the HeLa cells, but F protein was not efficiently enough to induce cell fusion in the co-expression system. |
|
|
|
Key words: NDV；M gene；NP gene；F gene；HN gene；eukaryotic expression;virus-like particles |
|
|
|
|
|
《中国兽医科学》 ©
版权所有
Chinese Veterinary Science
zgsykx@zgsykx.com |
