Selecting The Specific Genes Which Associated To F Protein Of HCV By Yeast Two-Hybridization And The Expression Of F

Yet, it was not until 1989 that the first sequences of hepatitis C virus (HCV) were reported.HCV's highly infection and serious damage on liver tissue call the scientists'attention. After many years hardwork, the life cycle and its gene products have been discovered .They pave the way for uncover the pathological process and effective therapies and medicines in the future. In addition, HCV is not clear completely. In 2001, a research group from University of Southern California reported the discovery of a novel HCV protein which is 16-17kd synthesized by ribosomal frameshift.And this protein has some overlap sequences with core gene and existed in many subtypies.The exact function of F is presently unknown. Some research groups reported that it can repress p21 expression.HCV F Protein comes from an alternate reading frame (AFR) that overlaps the core gene. We now use yeast hybrid system to select the specific genes which could combine to F protein so that we may predict the function of F on the base of these genes.Firstly, we reconstruct the new clone of PGBKT7-F as a bait,then transfer into the yeast AH109, After that using the liver cDNA library to associate to the reconstruction .We could select the yeast clones and test them by X-a-gal. Then extracting the plasmid of the yeasts and transfer them electrically into the Ecoli TOP10.Finally aligning the sequences on Gen Bank and decide the specific genes which may associate with the F. So far we have got 5 known genes: APOH, C8B, HP, RBP4, EEF2.Besides, F were expressed respectively in the Escherichia coli and T293 Cells to get expression information of the F protein.Finally, we use bioinformatics to analyze the sequence of F and predict epitopes and secondary structure of F.