Construction of plasmid coexpressing hepatitis B surface antigen and granulocyte macrophage-colony stimulating factor / 重庆医科大学学报

ABSTRACT

Objective:To construct plasmid coexpressing GM-CSF and HBsAg,enhance hepatitis B DNA vaccines antivirus effect.Methods:(1)The HBsAg-encoding fragment and GM-CSF-encoding fragment were created by PCR amplification from pEcob6 and pCD-hGM-CSF respectively,then were cloned into plasmid pcDNA3.1(+),pcDNA3.1-S and pcDNA3.1-GM-CSF(without stop codon)were constructed.(2)HBsAg DNA fragment (with stop codon) was amplified by PCR,then was subcloned into plasmid pcDNA3.1-GM-CSF.Results:Recombinant pcDNA3.1-S and pcDNA3.1-GM-CSF-S were confirmed by using restriction enzymes and DNA sequencing.HBsAg was detected in the lysates and supernatants from cells transfected with pcDNA3.1-S and in the lysates from cells transfected with pcDNA3.1-GM-CSF-S.Conclusion:Two eukaryotic expression plasmids were constructed successfully and could express HBsAg.