Open AccessAn inducible expression system permitting the efficient purification of a recombinant antigen from Mycobacterium smegmatis
Author(s) -
Triccas James A.,
Parish Tanya,
Britton Warwick J.,
Gicquel Brigitte
Publication year - 1998
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1998.tb13221.x
Subject(s) - mycobacterium smegmatis , recombinant dna , flag tag , biology , mycobacterium , immunogenicity , expression vector , microbiology and biotechnology , antigen , mycobacterium leprae , fusion protein , bacteria , biochemistry , gene , mycobacterium tuberculosis , genetics , medicine , tuberculosis , leprosy , pathology , immunology
A novel expression vector utilising the highly inducible acetamidase promoter of Mycobacterium smegmatis was constructed. High‐level induction of a model antigen, the Mycobacterium leprae 35 kDa protein, was demonstrated in recombinant M. smegmatis grown in the presence of the acetamidase inducer acetamide. The recombinant protein could be simply and efficiently purified from the bacterial sonicate by virtue of a C‐terminal 6‐histidine tag, demonstrating that this purification strategy can be used for the mycobacteria. The histidine tag had no apparent effect on the protein conformation or immunogenicity, suggesting that the vector described may prove useful for the purification of native‐like recombinant mycobacterial proteins from fast‐growing mycobacterial hosts.