[Prokaryotic expression and characterization of the Rv2994 gene from Mycobacterium tuberculosis].
OBJECTIVE
To construct a prokaryotic expression vector bearing Rv2994 gene from Mycobacterium tuberculosis and provide materials for investigating the function of the gene.


METHODS
The Rv2994 gene was amplified by Polymerase Chain Reaction from Mycobacterium tuberculosis H37Rv strain and cloned into prokaryotic expression vector pGEX-1lamdaXT. The recombinant plasmid pGEX-2994 was sequenced and transformed into E. coli JM109 to be induced with IPTG and expressed the 73kDa fusion protein GST-Rv2994. It's antigenicity was confirmed by Western blotting. The expression product was purified and immunized the new Zealand rabbits.


RESULTS
The Rv2994 gene was amplified accurately from the genome DNA of H37Rv. A recombinant fused expression vector pGEX-Rv2994 was constructed and GST-Rv2994 protein was purifiel to immunize New Zealand rabbit.


CONCLUSIONS
The prokaryotic expression vector pGEX-2994 was constructed, and the 73kDa fusion protein GST-Rv2994 was expressed and purified successfully. It provided the basis for the further study of the Rv2994 gene.
Zhao
Ming-cai
MC
Infection and Immunity Unit, West China Center of Medical Sciences, Sichuan University, Chengdu 610041, China.
Bao
Lang
L
Wu
Yue-han
YH
Zhang
Hui-dong
HD
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D009169:Mycobacterium tuberculosis / Q000235:genetics* / Q000378:metabolism; D011817:Rabbits; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D011993:Recombinant Fusion Proteins