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中文摘要：
以结核分枝杆菌H37Rv株的基因组作为模板，将Ag85B和ESAT6编码基因进行PCR扩增，然后采用基因剪接重叠扩增PCR法（gene SOEing）将其通过疏水甘氨酸接头（GGIGIAPG）连接融合，定向克隆至质粒Pvax1中，构建结核分枝杆菌Ag85B-ESAT6融合抗原的真核表达质粒Pvax1／AE．经单双限制性内切酶图谱、PCR产物及DNA测序分析等多种方法鉴定，证实Pvax1／AE真核表达质粒构建成功．为进一步研究其结核核酸疫苗原型的免疫保护效果奠定了基础．
英文摘要：
To construct eukaryotic expression vector expressing Mycobacterium tuberculosis fusion antigen Ag85B- ESAT6, the genes respectively encoding Ag85B and ESAT6 protein were amplified by polymerase chain reaction （PCR） based on genome of MTB H37Rv strain. Then by gene SOEing method, fusion gene encoding Ag85B-ESAT6 protein was linked with the Iinker（GGIGIAPG）. The fusion gene was cloned into pVAX1 . Using single and double restriction endonuclease digestion, PCR and DNA sequencing, it was confirmed that the recombinant eukaryotic expression plasmid （pVAX1/AE） had been successfully constructed. The result serves as a prototype of nucleic acid vaccine for further studies on its immunity effectiveness against MTB.
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