含禽流感病毒血凝素蛋白VSV假病毒制备

赖天莹; 李永刚; 单颖

doi:10.11847/zgggws1121915

含禽流感病毒血凝素蛋白VSV假病毒制备

Preparation of a pseudo vesicular stomatitis virus containing avian influenza virus hemagglutinin protein

摘要

摘要:
目的构建含有H6N2型禽流感病毒血凝素蛋白（HA）的疱疹口炎病毒（VSV）。
方法以H6N2亚型禽流感病毒互补脱氧核糖核酸（cDNA）为模板，扩增HA基因，并与pcDNA3.1 + 载体连接；使用VSVΔG*-G病毒感染制备VSVΔG*-HA假病毒，并对其血凝活性进行测定。
结果成功构建重组表达载体pcDNA3.1HA；HA蛋白表达于293T细胞；荧光显微镜证实VSVΔG*-HA假病毒包装成功；活性试验显示VSVΔG*-HA假病毒具有与禽流感病毒相似的血凝活性。
结论含有H6N2型禽流感病毒HA蛋白的VSV假病毒安全、可靠，为进一步对禽流感病毒受体等研究及筛选抗流感病毒的药物提供了技术支持和思路。

Abstract:
Objective To construct a vesicular stomatitis virus (VSV) containing hemagglutinin protein (HA) of H6N2 avian influenza virus.
Methods The H6N2 subtype avian influenza virus complementary deoxyribonucleic acid (cDNA) was used as a template to amplify the HA gene and was ligated to the pcDNA3.1 + vector. VSVΔG*-G virus infection was used to prepare VSVΔG*-HA pseudo virus. The VSVΔG*-HA virus of the recombinant gene was assayed and its hemagglutination activity was measured.
Results The recombinant expression vector pcDNA3.1-HA was successfully constructed. The HA protein was expressed in 293T cells. Fluorescence microscopy confirmed the successful packaging of VSVΔG*-HA pseudo virus. Activity test showed that the VSVΔG*-HA pseudovirus has similar hemagglutination activity to that of avian influenza virus.
Conclusion The pseudo VSV containing H6N2 avian influenza virus HA protein is safe and reliable, and could provide a technical support and ideas for further research and screening of anti-influenza drugs for avian influenza virus receptors.

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