Cloning and sequential analysis of ESAT6 gene of Mycobacterium tuberculosis H37Ra

Objective: To clone and sequence the ESAT6 gene from Mycobacterium tuberculosis(MTB) H37Ra.Methods: The encoding gene of ESAT6 was amplified from MTB H37Ra genomic DNA using PCR technique.The amplified PCR product was subcloned into pTG19-T vector.The target gene was identified by PCR,single and double enzyme digestion,sequencing and sequence alignment analysis with BLAST software.Results: The ESAT6 gene was successfully cloned.The sequencing showed the fragment length was 288 bp,which was homology with the MTB H37Ra gene reported by GenBank.Conclusions: The coding gene of ESAT6 from MTB H37Ra is successfully cloned.