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Expression of Escherichia coli β‐galactosidase in Mycobacterium bovis BCG using an expression system isolated from Mycobacterium paratuberculosis which induced humoral and cellular immune responses
Author(s) -
Murray A.,
Winter N.,
Lagranderie M.,
Hill D. F.,
Rauzier J.,
Timm J.,
Leclerc C.,
Moriarty K. M.,
Gheorghiu M.,
Gicquel B.
Publication year - 1992
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1992.tb02201.x
Subject(s) - biology , mycobacterium bovis , mycobacterium smegmatis , shuttle vector , escherichia coli , microbiology and biotechnology , plasmid , insertion sequence , paratuberculosis , recombinant dna , expression vector , open reading frame , gene , mycobacterium , transposable element , vector (molecular biology) , mycobacterium tuberculosis , bacteria , genetics , peptide sequence , mutant , medicine , tuberculosis , pathology
Summary A promoter sequence, P an' , was isolated from Mycobacterium paratuberculosis and characterized. This promoter lies adjacent to, and outside, the 3’end of an IS 900 insertion element. IS 900 contains an open reading frame, ORF2, on the complementary strand which codes for the putative transposase of this insertion sequence. A DNA fragment containing P AN and part of ORF2 was fused to the lacZ gene and inserted into the replicative shuttle vector pRR3. Mycobacterium smegmatis and Mycobacterium bovis BCG (BCG) transformed with this plasmid exhibited β‐galactosidase activity. However, lacZ was only expressed in Escherichia coli under the control of P an , when ORF2 was deleted. Immunization of mice with the recombinant M. bovis BCG expressing lacZ resulted in the induction of a high humoral and cellular response directed against β‐galactosidase. The P an ‐ORF2 expression system may prove to be particularly useful for cloning and expression of heterologous genes in the BCG vaccine strain.