Vero E6细胞酵母双杂交cDNA文库的构建及鉴定

Abstract

Abstract: To seek out proteins which interact with structural proteins of porcine epidemic diarrhea virus （PEDV） in Vero E6 cells and study the mechanisms of viral infection.We created a Vero E6 cDNA yeast hybrid library. The total RNA of Vero E6 was extracted using Trizol method, and double strands cDNA was synthesized by SMART technique. We created a Vero E6 cDNA library by using homologous recombination in yeast. The results showed that the titer of cDNA library was 9.7×10⁸ CFU with the average of inserted fragments about 1.6 kb, and the recombination rate was 87%. These data indicated that the yeast two-hybrid cDNA library of Vero E6 was successfully constructed and might be useful for screening the host proteins interacting with structure protein of PEDV.

Key words: porcine epidemic diarrhea virus （PEDV）; Vero E6 cell; cDNA library; SMART technique; yeast two-hybrid

CHI Yan-bin，SHI Da，ZHU Qing-he，CHEN Jian-fei，SHI Hong-yan，ZHANG Xin，LI Chang-long，HAN Xiao，LIU Ning，ZHAO Xin，FENG Li. Construction and Characterization of the Yeast Two-hybrid cDNA Library of Vero E6 Cells[J]. .

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Construction and Characterization of the Yeast Two-hybrid cDNA Library of Vero E6 Cells

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