Construction of hantavirus Z37 envelope glycoprotein genes G1 and G2 recombinant and its expression in eukaryocyte / 中华传染病杂志

ABSTRACT

Objective To construct hantavirus Z37 envelope glycoprotein genes G1 and G2 recombinant and express in eukaryocyte. Methods Using three pairs of primers based on hantavirus Z37 M gene sequences. PCR products F1, F2 and G2 were obtained by PCR from the Plasmid pGEMZ37(containing partial cDNA of G1) and Plasmid pCUMZ37(containing partial cDNA of G1 and whole cDNA of G2). G1 PCR products were also obtained by fusing F1 and F2 PCR products. The G1 and G2 PCR products were then digested by BamHⅠ and XhoⅠ and cloned into the corresponding sites of expression vector pcDNA3.1 respectively. The recombinants pcDNA3.1 G1 and pcDNA3.1 G2 were identified by digestion with endonuclease BamHⅠ and XhoⅠ and confirmed by sequencing. After transfecting COS 7 cells by Calcium phosphate/DNA precipitating method, indirect immunofluorescence assay (IFA) was used to verify the transient expression of HV Z37 G1 or G2 protein in COS 7 cells. Results The recombinant expression plasmids pcDNA3.1 G1 and pcDNA3.1 G2 were constructed. After transfection with the above recombinant expression plasmids, specific antigens were detected within cells by IFA. Conclusions The reombinant expression plasmids pcDNA3.1 G1、pcDNA3.1 G2 were constructed successfully and expressed transiently in eukaryocyte.