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PMID: 3020007 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Cloning and expression of Mycobacterium bovis BCG DNA in "Streptomyces lividans".

Journal of bacteriology ·Vol. 168 ·No. 1 ·1986-10-00 ·Pages 72-80

Kieser T, Moss MT, Dale JW, Hopwood DA

Abstract

The ability of "Streptomyces lividans" to use the expression signals of genes from Mycobacterium bovis BCG was tested in vivo by using gene fusions. Random DNA fragments from M. bovis BCG were inserted into promoter-probe plasmids in Escherichia coli and in "S. lividans." Comparison with promoter activity detected with random DNA fragments from the respective hosts suggested that "S. lividans" efficiently utilizes a high proportion of mycobacterial promoters, whereas a smaller fraction are expressed, and expressed more weakly, in E. coli. M. bovis BCG DNA fragments were also inserted into the specially constructed translational fusion vector (pIJ688) in "S. lividans." pIJ688 contains the kanamycin phosphotransferase gene (neo) from transposon Tn5, truncated at its amino terminus, as the indicator. The results suggested that "S. lividans" uses M. bovis BCG translational signals almost as efficiently as its own signals. Moreover, several hybrid proteins with an M. bovis BCG-derived amino terminus seemed to be reasonably stable in "S. lividans." These experiments indicate that "S. lividans" may be a suitable host for the expression of Mycobacterium leprae and Mycobacterium tuberculosis genes from their own signals. This is a precondition for the expression of entire biosynthetic pathways, which could be valuable in the production of diagnostic and therapeutic agents. The vectors may also have wider applications for the analysis of gene expression in Streptomyces.

MeSH Terms
Cloning, Molecular DNA Transposable Elements Drug Resistance, Microbial Escherichia coli/genetics Genes, Bacterial Kanamycin/pharmacology Kanamycin Kinase Mycobacterium bovis/genetics Phosphotransferases/biosynthesis,genetics Promoter Regions, Genetic Protein Biosynthesis Streptomyces/drug effects,enzymology,genetics
Chemicals
DNA Transposable Elements Kanamycin Phosphotransferases Kanamycin Kinase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Kieser T
Moss M T
Dale J W
Hopwood D A
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31 references, click to expand
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Article Info
Journal
Journal of bacteriology
Abbr.
J Bacteriol
ISSN
0021-9193
DOI
10.1128/jb.168.1.72-80.1986
Published
1986-10-00
Pages
72-80
Language
English
Region
United States
NLM ID
2985120R
PMCID
PMC213421
Subset
IM
External Links
PubMed source DOI: 10.1128/jb.168.1.72-80.1986 PMC Full Text View journal details
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