细胞因子GM-CSF和结核杆菌Ag85A融合表达载体的构建与鉴定-东南大学学报医学版

细胞因子GM-CSF和结核杆菌Ag85A融合表达载体的构建与鉴定

单位：东南大学基础医学院,病原生物学与免疫学系,江苏,南京,210009

分类号：R392.2, Q782

出版年·卷·期（页码）：2003·22·第四期（232-235）

摘要：

目的:构建并鉴定细胞因子GM-CSF和结核杆菌Ag85A融合表达质粒,为研究新型抗结核杆菌DNA疫苗提供新的策略.方法:用PCR方法从pc-mGM-CSF质粒中扩增出GM-CSF,构建于pcDNA3.1质粒上,成为pcDNA3.1-GM-CSF.从结核杆菌H37Rv基因组中扩增出结核杆菌Ag85A基因序列并插入到质粒pcDNA3.1-GM-CSF上,将基因GM-CSF的3′端与基因Ag85A的5′端直接连接构建融合表达质粒pcGM85A.结果:经酶切鉴定和DNA测序证实重组质粒构建正确.结论:细胞因子GM-CSF和结核杆菌Ag85A融合表达载体构建成功,为研制新型结核病基因疫苗奠定了基础.

Objective To construct and identify an eukaryotic expression plasmid containing GM?CSF and TB Ag85A.Methods GM?CSF was amplified from plasmid pc-mGM?CSF by PCR and cloned directly into pcDNA3.1 vector(GM?CSF gene without stop codon).By PCR,the gene of Ag85A was amplified from genome of mycobacterium tuberculosis H 37Rv and cloned directly into recombinant pcDNA3.1?GM?CSF and pcGM85A was finally formed.Result The eukaryotic expression recombinant plasmid containing GM?CSF and mycobacterium tuberculosis Ag85A has been constructed correctly by identification of restriction enzyme digesting and DNA sequencing confirmation.Conclusion An eukaryotic expression recombinant plasmid pcGM85A has been constructed successfully,which is an access to further researches on the development of new type anti-tuberculosis vaccine.

参考文献：

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