Nucleotide sequence of the streptothricin acetyltransferase gene from Streptomyces lavendulae and its expression in heterologous hosts
- 1 May 1987
- journal article
- research article
- Published by American Society for Microbiology in Journal of Bacteriology
- Vol. 169 (5), 1929-1937
- https://doi.org/10.1128/jb.169.5.1929-1937.1987
Abstract
The nucleotide sequence of the streptothricin acetyltransferase (STAT) gene from streptothricin-producing Streptomyces lavendulae predicts a 189-amino-acid protein of molecular weight 20,000, which is consistent with that determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the purified enzyme. The amino acid composition and the NH2-terminal sequence determined by using the purified protein are in good agreement with those predicted from the nucleotide sequence, except for the absence of the NH2-terminal methionine in the mature protein. High-resolution S1 nuclease protection mapping suggests that transcription initiates at or near the adenine residue which is the first position of the translational initiation triplet (AUG) of STAT. Another open reading frame located just upstream of the STAT gene was detected and contains a region bearing a strong resemblance to DNA-binding domains which are conserved in known DNA-binding proteins. By addition of promoter signals and a synthetic ribosome-binding (Shine-Dalgarno) sequence at an appropriate position upstream of the STAT translational start codon, the STAT gene confers streptothricin resistance on Escherichia coli and Bacillus subtilis. The STAT coding sequence with both the promoter of a B. subtilis cellulase gene and a synthetic Shine-Dalgarno sequence was functionally expressed in Streptomyces lividans, which suggests that the addition of an artificial leader upstream of the translational initiation codon (AUG) does not significantly influence the translation of STAT.This publication has 46 references indexed in Scilit:
- Construction and characterisation of a series of multi-copy promoter-probe plasmid vectors for Streptomyces using the aminoglycoside phosphotransferase gene from Tn5 as indicatorMolecular Genetics and Genomics, 1986
- Genetic and Biochemical Characterization of the red Gene Cluster of Streptomyces coelicolor A3(2)Microbiology, 1985
- RNA polymerase heterogeneity in Streptomyces coelicolorNature, 1985
- N-methyl transferase of Streptomyces erythraeus that confers resistance to the macrolidelincosamide-streptogramin B antibiotics: amino acid sequence and its homology to cognate R-factor enzymes from pathogenic bacilli and cocciGene, 1984
- Molecular cloning of the whole biosynthetic pathway of a Streptomyces antibiotic and its expression in a heterologous hostNature, 1984
- Promoter recognition by sigma-37 RNA polymerase from Bacillus subtilisJournal of Molecular Biology, 1984
- Cloning and Expression of the Tyrosinase Gene from Streptomyces antibioticus in Streptomyces lividansMicrobiology, 1983
- Physical analysis of antibiotic-resistance genes from Streptomyces and their use in vector constructionGene, 1982
- Homology among DNA-binding proteins suggests use of a conserved super-secondary structureNature, 1982
- Fate of transforming DNA following uptake by competent Bacillus subtilis: I. Formation and properties of the donor-recipient complexJournal of Molecular Biology, 1971