Large Amplification of a 35-kb DNA Fragment Carrying Two Penicillin Biosynthetic Genes in High Penicillin Producing Strains of Penicillium Chrysogenum

Curr Genet. 1989 Dec;16(5-6):453-9. doi: 10.1007/BF00340725.

Abstract

The isopenicillin N synthase (pcbC) and acyl-CoA:6-APA acyltransferase (penDE) genes of Penicillium chrysogenum were located in a 19.5-kb DNA fragment that had been previously cloned in phage vector EMBL3. This 19.5-kb DNA fragment was mapped with several endonucleases, and the pcbC and penDE genes were located by hybridization with probes corresponding to internal fragments of each gene. A low penicillin producing strain (P. chrysogenum Wis 54-1255) and two high producing strains (AS-P-78 and P2) showed hybridizing fragments of identical sizes in their chromosomes. Dot-blot hybridization of serial dilutions of the total DNA of the three strains showed that the intensity of all the hybridizing bands was much higher in strains AS-P-78 and P2 than in Wis 54-1255. Hybridization of total DNA digestions with probes corresponding to fragments which mapped upstream or downstream of the pcbC-penDE region revealed that a fragment of at least 35 kb DNA has been amplified 9 to 14 fold in the high penicillin producing strains. The amplified region did not include the previously cloned pyrG gene that encodes OMP-decarboxylase, an enzyme involved in pyrimidine biosynthesis.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acyltransferases / genetics*
  • Blotting, Southern
  • DNA, Fungal / genetics*
  • Enzymes / genetics*
  • Gene Amplification*
  • Genes, Fungal
  • Oxidoreductases*
  • Penicillin-Binding Proteins*
  • Penicillins / biosynthesis*
  • Penicillium / genetics*
  • Penicillium chrysogenum / genetics*
  • Penicillium chrysogenum / metabolism
  • Restriction Mapping

Substances

  • DNA, Fungal
  • Enzymes
  • Penicillin-Binding Proteins
  • Penicillins
  • Oxidoreductases
  • isopenicillin N synthetase
  • Acyltransferases
  • acyl-CoA-6-aminopenicillanic acid acyltransferase