Cloning and characterization of a rhamnogalacturonan hydrolase gene from Botrytis cinerea

Biochem Mol Biol Int. 1997 Nov;43(4):823-38. doi: 10.1080/15216549700204641.

Abstract

Rhamnogalacturonan hydrolase (Rgase A) cleaves alpha 1--2 linkages between rhamnosyl and galacturonosyl residues in pectin. A 1.9 kb RGase A cDNA clone (BCRHGA) was isolated from a B. cinerea cDNA library using a PCR-amplified Aspergillus aculeatus RGase A probe. It's 1.7 kb open reading frame had 62% identity at the amino acid level with A. aculeatus RGase A. Northern blots of B. cinerea total RNA probed with BCRHGA revealed a 2 kb band, suggesting the cDNA clone is full or nearly-full length. To determine mRNA expression of the gene, B. cinerea was grown in media containing 0.5% apple pectin, 0.5% rhamnogalacturonan-I and 1% glucose carbon sources. Northern analysis revealed the BCRHGA gene was expressed on all carbon sources, but with different patterns of expression. B. cinerea RGase A appeared to be coded for by a single or low copy number gene based on Southern analysis.

MeSH terms

  • Amino Acid Sequence
  • Base Sequence
  • Blotting, Northern
  • Blotting, Southern
  • Cloning, Molecular
  • DNA, Complementary / chemistry
  • DNA, Complementary / isolation & purification
  • DNA, Fungal / analysis
  • Fungal Proteins / biosynthesis
  • Fungal Proteins / chemistry*
  • Fungal Proteins / genetics*
  • Gene Expression Regulation, Fungal
  • Genes, Fungal*
  • Glycoside Hydrolases / biosynthesis
  • Glycoside Hydrolases / chemistry*
  • Glycoside Hydrolases / genetics*
  • Mitosporic Fungi / enzymology*
  • Mitosporic Fungi / genetics*
  • Molecular Sequence Data
  • Sequence Analysis, DNA

Substances

  • DNA, Complementary
  • DNA, Fungal
  • Fungal Proteins
  • Glycoside Hydrolases
  • rhamnogalacturonase A

Associated data

  • GENBANK/U62397