OL-049 Screening of proteins binding to HCV NS3 protein from human pancreas cDNA library by yeast two-hybrid

C.C. Hu, J.Q. Zhang, X.C. Wang, G.L. Li, Q. Wang, J. Cheng
2008 International Journal of Infectious Diseases  
Objective: To screen proteins of human pancreas cDNA library interacting with HCV NS3 protein. Methods: The library was amplifed, purified and evaluated, and then the puried library plasmids were transformed into yeast strain Y187. The reconstructed plasmid pGBKT7-NS3 was transformed into yeast strain AH109 and screened on the nutrient deficiency medium SD/-Trp. The transformed AH109 mated with Y187 containing the library plasmid. The diploid yeast cells were plated on nutrient deficiency
more » ... nt deficiency medium SD/-Trp/-Leu/-His/-Ade and SD/-Trp/-Leu/-His/-Ade containing X-a-gal for selecting. The plasmids in diploid yeast cells were extracted and electrotransformed into E. coli DH5a. The plasmids in DH5a were extracted, sequenced and analyzed by bioinformatic methods. Results: Eight proteins interacting with HCV NS3 were found. Conclusions: These results show that HCV NS3 protein may be related with metabolism of glucose and lipid.
doi:10.1016/s1201-9712(09)60146-6 fatcat:wlmlfa4ugzbrnc446v2kwf7x5i