Cloning and expression of Mycobacterium bovis BCG DNA in "Streptomyces lividans".
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.
D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D004352:Drug Resistance, Microbial; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D007612:Kanamycin / Q000494:pharmacology; D019868:Kanamycin Kinase; D009163:Mycobacterium bovis / Q000235:genetics*; D010770:Phosphotransferases / Q000096:biosynthesis / Q000235:genetics; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D013302:Streptomyces / Q000187:drug effects / Q000201:enzymology / Q000235:genetics*
D004251:DNA Transposable Elements; D007612:Kanamycin; D010770:Phosphotransferases; D019868:Kanamycin Kinase