CONSTRUCTION OF GENE LIBRARY CONTAINING GROUP A ROTAVIRUS VP7 GENE

[Objective]To isolate and culture Group A rotavirus, then the outer capsid protein VP7 gene was cloned and transformed into pMD18-T vector and succeeded conducting cDNA gene library, so as to lay a foundation for the construction of genetic engineering vaccine against rotavirus. [Methods] Through extracted the total Group A rotavirus RNA and amplified by RT-PCR, the VP7 gene fragment was isolated and recovered, and then transformed into pMD18-T. After VP7 gene was extracted from the plasmid, it was detected by 1% agarose gel electrophoresis. [Results] The results of total RNA of Group A rotavirus were as following: the 5s, 18s and 28s band were explicit, the integration of VP7 gene was confirmed. The VP7 gene was transformed into pMD18-T. After screening by blue and white plague, the recombinant DNA (vector and the VP7 gene) was selected. [Conclusion] Through culturing of Group A rotavirus, amplifying VP7 gene, recovering DNA fragments, connecting DNA fragments, transforming and detecting DNA fragments, the VP7 gene has been successfully transformed into pMD18-T vector, succeeded conducting cDNA gene library, so as to lay a foundation for the further development of genetic engineering vaccine against rotavirus.