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2006•Letters in BiotechnologyRequires access

Cloning of esat6-cfp10 Fusion Gene and its Expression in Mycobacterium smegmatis

Zhang Haia

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Abstract

Objective To construct a recombinant plasmid carrying esat6 and cfp10 gene and express this fusion protein in Mycobacterium smegmatis. Methods: cfp10 gene was amplified by PCR with specific primer from genome of Mycobacterium tuberculosisMTB H37Rv. cfp10 was inserted into the cloning vector pGEM-T-easy with esat6 gene. Then esat6-cfp10 digested by restriction enzyme it was cloned to expressing vector pDE22 shuttle plasmid and expressed in Mycobacterium smegmatis. Results SDS-PAGE showed that a 23 000 fusion protein was expressed. Western blot suggested that this fusion protein has a good immunogenicity. Conclusion: The recombinant mycobacterium smegmatis expressed ESAT6-CFP10 fusion protein was obtained providing bases for preventing tuberculosis.

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What this paper is about

Objective To construct a recombinant plasmid carrying esat6 and cfp10 gene and express this fusion protein in Mycobacterium smegmatis. Methods: cfp10 gene was amplified by PCR with specific primer from genome of Mycobacterium tuberculosisMTB H37Rv. cfp10 was inserted into the cloning vector pGEM-T-easy with esat6 gene. Then esat6-cfp10 digested by restriction enzyme it was cloned to expressing vector pDE22 shuttle plasmid and expressed in Mycobacterium smegmatis. Results SDS-PAGE showed that a 23 000 fusion protein was expressed. Western blot suggested that this fusion protein has a good immunogenicity. Conclusion: The recombinant mycobacterium smegmatis expressed ESAT6-CFP10 fusion protein was obtained providing bases for preventing tuberculosis.

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Available abstract

Objective To construct a recombinant plasmid carrying esat6 and cfp10 gene and express this fusion protein in Mycobacterium smegmatis. Methods: cfp10 gene was amplified by PCR with specific primer from genome of Mycobacterium tuberculosisMTB H37Rv. cfp10 was inserted into the cloning vector pGEM-T-easy with esat6 gene. Then esat6-cfp10 digested by restriction enzyme it was cloned to expressing vector pDE22 shuttle plasmid and expressed in Mycobacterium smegmatis. Results SDS-PAGE showed that a 23 000 fusion protein was expressed. Western blot suggested that this fusion protein has a good immunogenicity. Conclusion: The recombinant mycobacterium smegmatis expressed ESAT6-CFP10 fusion protein was obtained providing bases for preventing tuberculosis.

Key concepts: Mycobacterium smegmatis, Shuttle vector, Fusion protein, Plasmid, Mycobacterium tuberculosis, Restriction enzyme, Recombinant DNA, Biology

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Cloning of esat6-cfp10 Fusion Gene and its Expression in Mycobacterium smegmatis — Research Paper | ScholarLens