[H5N1流感病毒M2和NA基因表达载体的构建]

[Construction of vectors expressing M2 and NA genes of H5N1 influenza virus].

作者信息

Guo Jian-Qiang, Yao Li-Hong, Chen Ai-Jun, Liu Xiao-Yu, Fu Jin-Qi, Xu Peng-Wei, Zhang Zhi-Qing

机构信息

State Key Laboratory for Molecular Virology and Genetic Engineering, Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052, China.

出版信息

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2011 Jun;25(3):167-9.

PMID:21977581

Abstract

OBJECTIVE

To construct vectors expressing M2 and NA genes of H5N1 influenza virus.

METHODS

Based on the human H5N1 avian influenza virus (A/Anhui/1/2005) isolated in china, M2 and NA genes were amplified by PCR. M2 or NA gene was subcloned into pStar vector to construct recombinant pStar-M2/, pStar-/M2, pStar-NA/and pStar-NA/. Furthermore, both of the M2 and NA genes were subcloned into pStar to construct two genes co-expressing recombinant pStar-M2/NA and pStar-NA/M2. Expression of the genes were detected by IFA after transfection of 293 cells with the recombinant plasmids.

RESULTS

Recombinant plasmids were constructed and identified by restriction endonuclease digestion. Expression of the genes cloned into the recombinant plasmids was confirmed by IFA.

CONCLUSION

Recombinant plasmids expressing M2 and/or NA genes of H5N1 influenza virus were constructed, which provided basis for development of influenza DNA vaccine.

摘要

目的

构建表达H5N1流感病毒M2和NA基因的载体。

方法

以中国分离的人源H5N1禽流感病毒（A/Anhui/1/2005）为基础，通过PCR扩增M2和NA基因。将M2或NA基因亚克隆到pStar载体中，构建重组体pStar-M2/、pStar-/M2、pStar-NA/和pStar-NA/。此外，将M2和NA基因都亚克隆到pStar中，构建双基因共表达重组体pStar-M2/NA和pStar-NA/M2。用重组质粒转染293细胞后，通过免疫荧光分析（IFA）检测基因的表达。