[新城疫病毒LaSota株基因P和NP在两种不同细胞中的瞬时表达与鉴定]

[Transient expression and identification of gene P and NP of NDV LaSota strain in two different cells].

作者信息

Zhou Yuan, Jia Run-Qing, Teng Zhi-Ping, Zhang Xiao-Mei, Zeng Yi

机构信息

The College of Life Science and Bio-engineering, Beijing University of Technology, Beijing 100124, China.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2010 Feb;24(1):62-4.

PMID:20848855

Abstract

OBJECTIVE

To Construction of P and NP genes eukaryotic expression vectors of Newcastle Disease Virus LaSota strain,study its reverse genetics and functional genome of NDV.

METHODS

P, NP genes were amplified and cloned into pGEM-T easy vector and then subcloned into pcDNA3.1 (+) expression vector respectively, the recombinant plasmids were named pcDNA3.1 (+)-P and pcDNA3.1 (+)-NP, Recombinant plasmids were transfected into 293 and BHK-21 cells respectively and were detected using IE and Western blot analysis.

RESULTS

Expression of P, NP genes were detected and confirmed by the IE and WB analysis.

CONCLUSION

The recombinant eukaryotic plasmids pcDNA3. 1(+)-P, pcDNA3.1 (+)-NP were expressed in 293 and BHK-21 cells successfully. This research may be helpful for further study of reverse genetics and functional genome of NDV.

摘要

目的

构建新城疫病毒LaSota株P和NP基因的真核表达载体，研究其反向遗传学及新城疫病毒功能基因组。

方法

扩增P、NP基因并克隆至pGEM-T easy载体，然后分别亚克隆至pcDNA3.1(+)表达载体，重组质粒分别命名为pcDNA3.1(+)-P和pcDNA3.1(+)-NP，将重组质粒分别转染至293和BHK-21细胞，并用免疫荧光（IE）和蛋白质免疫印迹（Western blot）分析进行检测。