Construction and identification of shuttle plasmid of rBCG

VernacularTitle:GCA融合基因重组BCG穿梭质粒的构建与鉴定
Author: Shangdan NIE ¹ ; Weiping LIANG ; Qingjie XUE
Author Information

1. 济宁医学院法医学与医学检验学院
Keywords: BCG vaccine; Genes; DNA,recombinant
From: Chinese Journal of Experimental and Clinical Virology 2015;29(4):361-363
CountryChina
Language:Chinese
Abstract: Objective To construct a secretory recombinant Bacillus Calmette-Guerin (BCG)plasmid with Ag85B fused LMP2A and GM-CSF genes.Methods BCG-Ag85B signal sequence and GCA gene were amplified from the genomes of BCG and GCA by PCR respectively,BCG-Ag85B signal sequence was cloned in E.coli-BCG shuttle-vector pMV261 to obtain pMVS.Recombinant plasmid pMVGCA was constructed by inserting the GCA gene into pMVS.Results The cloned genes BCG-Ag85B and GCA were correctly inserted into the plasmid pMV261,which had been identified by restriction endonuclease digestion and PCR amplification of GCA and gene sequencing respectively.Conclusion The plasmid pMV261-GCA was expected to secretively express GCA in BCG.This research had provided the possibility of further researches on the development of new EB virus cancer vaccine.