[以Ad41为载体表达人轮状病毒VP6基因的重组腺病毒的遗传稳定性]
[The genetic stability of recombinant adenovirus expressing human rotavirus VP6 gene which used Ad41 as vector].
作者信息
Wang Min, Chen Duo-Ling, Song Jing-Dong, Qu Jian-Guo, Lu Zhuo-Zhuang, Zhao Wei-Ming, Hong Tao
机构信息
Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing, China.
出版信息
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2012 Dec;26(6):422-4.
OBJECTIVE
To investigate the genetic stability of non-replicating recombinant adenovirus which used Ad41 as vector and could express VP6 gene of group A rotavirus during continous passage, in order to develop the vaccine of rotavirus.
METHODS
The recombinant adenovirus rvAd41-VP6 (o) was prepared by our laboratory early, it then was continuously propagated on 293TE7 cells for 14 passages. After that samples of the infected cells were collected at every 2 passages for the detection of the integration of the VP6 gene by PCR, and the expression of the target protein was detected by Western Blot analysis.
RESULTS
Analysis by PCR revealed that, there was stable integration of specific VP6 gene in the rvAd41-VP6 (o), Western Blot analysis confirmed that rvAd41-VP6 (o) could stably expressed the group-specific antigen structural protein VP6 (o), and it had preferable genetic stability.
CONCLUSION
The recombinant adenovirus rvAd41-VP6 (o) which could stably express the VP6 (o) gene had favorable biological property in vitro, and it has provided a basis for further research of animal immunization.
目的
研究以Ad41为载体、可表达A组轮状病毒VP6基因的非复制型重组腺病毒在连续传代过程中的遗传稳定性，以研制轮状病毒疫苗。
方法
本实验室前期制备重组腺病毒rvAd41-VP6（o），然后在293TE7细胞上连续传代14次。之后每传2代收集感染细胞样本，通过PCR检测VP6基因的整合情况，通过蛋白质免疫印迹分析检测目的蛋白的表达情况。
结果
PCR分析显示，rvAd41-VP6（o）中特异性VP6基因稳定整合；蛋白质免疫印迹分析证实rvAd41-VP6（o）可稳定表达组特异性抗原结构蛋白VP6（o），且具有较好的遗传稳定性。
结论
可稳定表达VP6（o）基因的重组腺病毒rvAd41-VP6（o）在体外具有良好的生物学特性，为进一步开展动物免疫研究奠定了基础。
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