Wei Qiang, Jiang Xiu-li, Wang Jian-wei, Hong Tao
Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052, China.
Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi. 2004 Mar;18(1):12-5.
To express main neutralization antigen VP7 of human rotavirus serotype G2 and G3 by recombinant adenoviruses on the basis of previous investigation of the prevalence of rotavirus in China.
On the basis of successfully expression of human rotavirus protein G1VP7 by recombinant adenovirus vector, the authors constructed some potent recombinant adenovirus strains encoding rotavirus G2 VP7 and G3 VP7 genes which belong to the main rotavirus isolates 97S43 and 97S48.
Replication defective recombinant adenoviruses expressing human rotavirus serotype G2 and G3 VP7 genes, named as rvAdG2VP7 and rvAdG3VP7 were successfully constructed. VP7 genes integrated into the viral genome were identified by PCR and Southern blot assay, and specific transcription were detected by RT-PCR in the 293 cells infected with recombinant adenoviruses. Expression of rotavirus VP7 proteins was demonstrated by Western blot assay.
The established recombinant adenoviruses expressing G2 and G3 serotype VP7s laid a significant basis for further animal experiments in the development of multivalent rotavirus vaccines against rotavirus infection.
基于此前对中国轮状病毒流行情况的调查，通过重组腺病毒表达人轮状病毒G2和G3型主要中和抗原VP7。
在重组腺病毒载体成功表达人轮状病毒蛋白G1VP7的基础上，构建了一些编码属于主要轮状病毒分离株97S43和97S48的轮状病毒G2 VP7和G3 VP7基因的高效重组腺病毒株。
成功构建了表达人轮状病毒G2和G3型VP7基因的复制缺陷型重组腺病毒，命名为rvAdG2VP7和rvAdG3VP7。通过PCR和Southern印迹分析鉴定了整合到病毒基因组中的VP7基因，并通过RT-PCR在感染重组腺病毒的293细胞中检测到特异性转录。通过Western印迹分析证实了轮状病毒VP7蛋白的表达。
所建立的表达G2和G3型VP7的重组腺病毒为进一步开展抗轮状病毒感染多价轮状病毒疫苗的动物实验奠定了重要基础。
