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PMID: 1537349 Published · ppublish English Comparative Study Journal Article Research Support, Non-U.S. Gov't

Cloning of the complete biosynthetic gene cluster for an aminonucleoside antibiotic, puromycin, and its regulated expression in heterologous hosts.

The EMBO journal ·Vol. 11 ·No. 2 ·1992-02-00 ·Pages 785-92

Lacalle RA, Tercero JA, Jiménez A

Abstract

Puromycin, produced by Streptomyces alboniger, is a member of the large group of aminonucleoside antibiotics. The genes pac and dmpM, encoding a puromycin N-acetyl transferase and an O-demethyl puromycin O-methyltransferase, respectively, are tightly linked in the DNA of S. alboniger. The entire set of genes encoding the puromycin biosynthesis pathway was cloned by screening a gene library from S. alboniger, raised in the low copy number cosmid pKC505, with a DNA fragment containing pac and dmpM. Puromycin was identified by biochemical and physicochemical methods, including 1H NMR, in the producing transformants. This pathway was located in a single DNA fragment of 15 kb which included the resistance, structural and regulatory genes and was expressed when introduced into two heterologous hosts Streptomyces lividans and Streptomyces griseofuscus. In addition to pac and dmpM, two other genes have been identified in the pur cluster: pacHY, which determines an N-acetylpuromycin hydrolase and prg1, whose deduced amino acid sequence is significantly similar to that of degT, a Bacillus stearothermophilus pleiotropic regulatory gene.

Related Genes
dmpM pac
MeSH Terms
Acetyltransferases/genetics,metabolism Amino Acid Sequence Cloning, Molecular Cosmids DNA, Bacterial/genetics,isolation & purification Escherichia coli/genetics Gene Library Genes, Bacterial Genetic Complementation Test Kinetics Methyltransferases/genetics,metabolism Molecular Sequence Data Multigene Family Open Reading Frames Plasmids Puromycin/biosynthesis,isolation & purification Restriction Mapping Sequence Homology, Nucleic Acid Streptomyces/enzymology,genetics,growth & development
Chemicals
DNA, Bacterial Puromycin Methyltransferases O-demethylpuromycin methyltransferase Acetyltransferases puromycin N-acetyltransferase
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Lacalle R A
Centro de Biología Molecular (CSIC and UAM), Universidad Autónoma, Madrid, Spain.
Tercero J A
Jiménez A
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1992-02-00
Pages
785-92
Language
English
Region
England
NLM ID
8208664
PMCID
PMC556512
Subset
IM
External Links
PubMed source PMC Full Text View journal details
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