猪小肠上皮细胞酵母双杂交cDNA文库的构建及鉴定

Abstract

Abstract: In order to study the molecular mechanism of invasion and virulence of the swine intestinal virus systematically,we constructed a porcine intestinal epithelial cells (IEC) cDNA yeast two-hybrid library. Taking advantage of RNeasy Min Kit,total cellular RNA was extracted from cultured porcine IEC,after reverse transcription,synthetise double stranded cDNA by the SMART technology,porcine IEC yeast cDNA library were constructed with homologous recombination. The results showed,library titer was 8.4×10⁸ CFU/mL,the inserted fragment of ds-cDNA was 0.5 to 2.0 kb,the average length up to 1.1 kb with a high library recombination rate. This library could be used for screening intestinal host proteins which were interacted with virus proteins,provide reference for the further study of the pathogenesis of the virus,and provide basement for the development of effective drug or vaccine.

Key words: porcine diarrhea virus; porcine intestinal epithelial cells; yeast two-hybrid; cDNA library

CLC Number:

XU Tian, SHI Da, CHEN Jian-fei, GU Feng-li, SHI Hong-yan, ZHANG Xin, FENG Li. Construction and Identification of Yeast Two-hybrid cDNA Library of Porcine Intestinal Epithelial Cells[J]. , 2014, 41(10): 33-37.

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Construction and Identification of Yeast Two-hybrid cDNA Library of Porcine Intestinal Epithelial Cells

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