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[Construction and identification of recombinant shuttle-plasmid with ESAT-6 from Mycobacterium tuberculosis]
Wei Chen1, Lang Bao, Yongen Xie
1Institute of Infection and Immunity, West China School of Preclinic and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Objective:
To construct a recombinant BCG secretively expressing ESAT-6 of Mycobacterium tuberculosis.
Methods:
alpha-antigen(alpha-Ag) signal sequence and esat-6 gene were amplified from the genome of Bacille Calmette-Guerin (BCG) and Mycobacterium tuberculosis by PCR respectively. esat-6 gene was cloned in E. coli-BCG shuttle-plasmid pMV261 to get pME. Then a new recombinant plasmid pSME was constructed by inserting BCG alpha-Ag signal sequence into pME.
Results:
The cloned genes alpha-Ag signal sequence and esat-6 were correctly inserted into the vector pMV261, which was confirmed by restriction endonuclease digestion and PCR amplification of pSME.
Conclusion:
pSME was expected to secretively express ESAT-6 of Mycobacterium tuberculosis in BCG. This study provides the possibility of further researches on the development of new anti-tuberculosis vaccine.