Cloning Of A Fusion Gene Containing Hgm-Csf Gene And Gene Encoding L Protein Of Hbv And Expression In Pichia Pastoris

Background/Aims: To construct the yeast expression vector containing cDNA sequence coding L protein of Hepatitis B Virus (HBV) and human granulocyte-macrophage colony stimulating factor (hGM-CSF), and to explore the method of secretory expression in Pichia Pastoris GS115 strain.Methods: We used pVAX1-L-hGM as template to amplify L-hGM gene by PCR, and then the PCR products were cloned into the yeast expression vector pPICZ alpha C with DNA recombination method. After linearized by Sac I, the recombinant plasmid pPICZ alpha C-L-hGM was transformed into Pichia Pastoris GS115 by electrophoresis. The expressing protein was induced by methanol. SDS-PAGE and Western blot were used to analyze the expression of protein.Results: DNA sequence analysis revealed that the constructed genes sequences were totally consistent with the Gene Bank reported. The results of SDS-PAGE and Western blot showed that the recombinant protein was induced by methanol and stably expressed in Pichia Pastoris.Conclusions: The successful construction of a recombinant yeast expression vector containing gene coding L protein of Hepatitis B virus and hGM-CSF gene, and expressed in Pichia Pastoris, of which lays a foundation for the further researches on a better protective HB vaccine.