Herpes simplex virus type 1 truncated glycoprotein B gene cloning and sequence identification

Objective To construct an eukaryotic expression plasmid of herpes simplex virus I(HSV-1)truncated glycoprotein B and to make the foundation for the construction of multi-valent DNA vaccine.Methods The encoding sequence of the truncated glycoprotein B was amplified from HSV-I SM44 DNA genome by means of polymerase chain response,incised by two different endonucleases,and then was directionally integrated into eukaryotic expression vector pcDNA3,and the recombinant vector pcDNA3-gBt was constructed,which was confirmed by the restriction endonuclease analysis and DNA sequencing.Results Electrophoresis of the products of double-enzyme digestion and analysis of the recombinant vector pcDNA3-gBt showed two bands,one was HSV-1 gBt gene(1.5 kb)and the other was linear pcDNA3(5.4 kb);DNA sequencing showed that the insertion orientation was right and the rate of homology was 99.5% compared with HSV-F of GeneBank.Conclusions The recombinant vector was constructed successfully,which would provide a theoretical basis to evaluate the possibility of the construction of multi-valent DNA vaccine for HSV-Ⅰ