Human hepatitis B virus polymerase expressed in Saccharomyces cerevisiae.docVIP

 PAGE \* MERGEFORMAT 14 Human hepatitis B virus polymerase expressed in Saccharomyces cerevisiae [Abstract] Objective To try to express in yeast human hepatitis B polymerase protein (HBV-Pol, and the team with a nickel-based protein particles were partially purified crude extract. Methods to full-length HBV-Pol gene was amplified by PCR and connected to High expression in Saccharomyces cerevisiae plasmid. transformants at 30 induced 3 d after 16 h, 4 to collect protein extracts and purified with nickel-based particles. detected by Western blot the expression of the results. The results re- The HBV-Pol present in this type of yeast protein extracts, this protein product may be Nickel-particle extraction. Conclusion The HBV-Pol in order to achieve a high level of expression of Sc in providing a reliable basis, but also easy to HBV- biochemical properties of Pol, and protein purification and antibody preparation further. [Keywords:] Hepatitis B virus; polymerase; Saccharomyces cerevisiae Abstract: Objective Attempt to express HBV-Pol in yeast expression system and then partially purify the rough extract with nickel-based particle. Methods Full length HBV-Pol gene was amplified and was introduced into a high effective expression plasmid of Saccharomyces cerevisiae (Sc. The transformant was cultured at 30 for 3 days, and then induction was carried out for another 16 hours at 30 , and then induction was carried out for another 16 hours at 30 . Cell lysis and purification were preformed at 4 . Target protein was detected in harvested rough extract with western blot. Results The recombinant HBV-Pol was present in rough extract of yeast and could be purified with nickel particle. Conclusions Results of this study prove that HBV-Pol can be expressed in Sc yeast and might facilitate the biochemical study, protein purification and antibody preparation of HBV-Pol. Keywords:: hepatitis B virus; polymerase; Saccharomyces cerevisiae Hepatitis B virus (Hepatit