CONSTRUCTION AND IDENTIFICATION OF EUKARYOTIC VECTOR EXPRESSING MYCOBACTERIUM TUBERCULOSIS FUSION ANTIGEN AG85B-ESAT6

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 linker(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.