【摘要】 目的构建分泌性表达IFN-α2a的卡介苗重组质粒。方法分别以卡介苗(BCG)和IFN-α2a cD-NA为模板,通过PCR扩增得到约117bp的BCG-Ag85B信号肽序列和495bp的IFN-α2a基因序列。将BCG-Ag85B信号肽序列与大肠杆菌-卡介苗穿梭表达载体pMV261重组,得到重组质粒pMS。再将IFN-α2a基因序列克隆至pMS中,得到重组质粒pMSIFN-α2a。结果质粒pMSIFN-α2a用双酶切和PCR扩增及测序鉴定证实,克隆基因BCG-Ag85B和IFN-α2a正确插入载体pMV261。结论重组质粒pMSIFN-α2a可望在BCG中分泌性表达细胞因子IFN-α2a,从而促进IFN-γ的释放,该质粒的构建成功为改造卡介苗、发展新型抗膀胱肿瘤疫苗奠定了基础。更多还原

【Abstract】 Objective To construct a recombinant Bacillus Calmette-Guérin(BCG) secreting human Inteferon(IFN)-α2a.Methods BCG Ag85B signal sequence and IFN-α2a gene were amplified from the genome of BCG and IFNα2a by PCR,respectively.BCG Ag85B signal sequence was cloned in E.Coli-BCG shuttle-vector pMV261 to get pMS.Then a new recombinant plasmid pMSIFN-α2a was constructed by inserting IFN-α2a gene into pMS.Results The cloned genes BCG Ag85B and IFN-α2a were correctly inserted into the vector pMV261,which was confirmed by restriction endonuclease digestion and PCR amplification of IFN-α2a and gene sequencing,respectively.Conclusions pMSIFN-α2a is expected to secretively express IFN-α2a of cytokine in BCG.This study provides the possibility of further researches on the development of new anti-bladder cancer vaccine.更多还原