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J. Biol. Chem., Vol. 265, Issue 27, 16358-16365, Sep, 1990
B Diez, S Gutierrez, JL Barredo, P van Solingen, LH van der Voort and JF Martin
Penicillium chrysogenum DNA fragments cloned in EMBL3 or cosmid vectors
from the upstream region of the pcbC-penDE cluster carry a gene (pcbAB)
that complemented the deficiency of alpha-aminoadipyl-cysteinyl-valine
synthetase of mutants npe5 and npe10, and restored penicillin production to
mutant npe5. A protein of about 250 kDa was observed in sodium dodecyl
sulfate-polyacrylamide gel electrophoresis gels of cell- free extracts of
complemented strains that was absent in the npe5 and npe10 mutants but
exists in the parental strain from which the mutants were obtained.
Transcriptional mapping studies showed the presence of one long transcript
of about 11.5 kilobases that hybridized with several probes internal to the
pcbAB gene, and two small transcripts of 1.15 kilobases that hybridized
with the pcbC or the penDE gene, respectively. The transcription initiation
and termination regions of the pcbAB gene were mapped by hybridization with
several small probes. The region has been completely sequenced. It includes
an open reading frame of 11,376 nucleotides that encodes a protein with a
deduced Mr of 425,971. Three repeated dominia were found in the
alpha-aminoadipyl- cysteinyl-valine synthetase which have high homology
with the gramicidin synthetase I and tyrocidine synthetase I. The pcbAB is
linked to the pcbC and penDE genes and is transcribed in the opposite
orientation to them.
The cluster of penicillin biosynthetic genes. Identification and characterization of the pcbAB gene encoding the alpha-aminoadipyl- cysteinyl-valine synthetase and linkage to the pcbC and penDE genes
Department of Ecology, Genetics and Microbiology, University of Leon, Spain.
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