caa a22 4500 465757553 CHVBK 20180323111851.0 cr unu---uuuuu 170327e19901201xx s 000 0 eng 10.1007/BF00327023 doi (NATIONALLICENCE)springer-10.1007/BF00327023 Gene disruption of the pcb AB gene encoding ACV synthetase in Cephalosporium acremonium [Elektronische Daten] [Jo Hoskins, Niamh O'Callaghan, Stephen Queener, Cathleen Cantwell, John Wood, Victor Chen, Paul Skatrud] Summary: Plasmid pPS 96 was used to disrupt the genomic region immediately upstream of pcbC in C. acremonium by homologous integration. Approximately 4% of the C. acremonium transformants obtained with pPS 96 were unable to produce beta-lactam antibiotics. All transformants obtained with other plasmids and isolates which had not been exposed to transforming DNA retained the ability to produce beta-lactams. Enzyme analysis showed that ACV synthetase activity was missing in the beta-lactam-minus pPS 96 transformants. Southern hybridization analysis confirmed the presence of multiple copies of pPS 96 in all beta-lactam-minus transformants analyzed. However, predictable alterations of the targeted region were not detected. Transformation of antibioticminus transformants with plasmid pZAZ4, carrying a wild-type copy of the region targeted for disruption, resulted in restoration of the ability to produce beta-lactams in greater than 80% of the transformants recovered. Location of the pcbAB gene upstream from pcbC was confirmed by comparing the amino acid sequence of internal peptides from purified ACV synthetase with that deduced from the DNA sequence of the region targeted for disruption. The direction of transcription of the pcbAB gene is opposite that of the pcbC gene. Further analysis of amino acid sequence data from ACV synthetase revealed regions of strong similarity with the peptide synthetases responsible for production of tyrocidine and gramicidin S in Bacillus brevis. Springer-Verlag, 1990 Cephalosporium acremonium nationallicence ACV synthetase nationallicence Gene disruption nationallicence pcb AB nationallicence Beta-lactam antibiotics nationallicence Hoskins Jo Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut O'Callaghan Niamh Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut Queener Stephen Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut Cantwell Cathleen Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut Wood John Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut Chen Victor Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut Skatrud Paul Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut Current Genetics Springer-Verlag 18/6(1990-12-01), 523-530 0172-8083 18:6<523 1990 18 294 https://doi.org/10.1007/BF00327023 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/BF00327023 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Hoskins Jo Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut NATIONALLICENCE 700 1- O'Callaghan Niamh Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut NATIONALLICENCE 700 1- Queener Stephen Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut NATIONALLICENCE 700 1- Cantwell Cathleen Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut NATIONALLICENCE 700 1- Wood John Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut NATIONALLICENCE 700 1- Chen Victor Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut NATIONALLICENCE 700 1- Skatrud Paul Lilly Research Laboratories, Lilly Corporate Center, 46285, Indianapolis, IN, USA aut NATIONALLICENCE 773 0- Current Genetics Springer-Verlag 18/6(1990-12-01), 523-530 0172-8083 18:6<523 1990 18 294 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer