title: Cloning and characterization of a rhamnogalacturonan hydrolase gene from Botrytis cinerea.
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.
authors.affiliation: Horticultural Crops Quality Laboratory, USDA, Agricultural Research Service, Beltsville, MD 20705, USA.
authors.forename: H J
authors.initials: HJ
authors.lastname: Chen
authors.forename: D L
authors.initials: DL
authors.lastname: Smith
authors.forename: D A
authors.initials: DA
authors.lastname: Starrett
authors.forename: D
authors.initials: D
authors.lastname: Zhou
authors.forename: M L
authors.initials: ML
authors.lastname: Tucker
authors.forename: T
authors.initials: T
authors.lastname: Solomos
authors.forename: K C
authors.initials: KC
authors.lastname: Gross
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004271:DNA, Fungal / Q000032:analysis; D005656:Fungal Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D006026:Glycoside Hydrolases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics*; D003904:Mitosporic Fungi / Q000201:enzymology* / Q000235:genetics*; D008969:Molecular Sequence Data; D017422:Sequence Analysis, DNA
chemical_list: D018076:DNA, Complementary; D004271:DNA, Fungal; D005656:Fungal Proteins; D006026:Glycoside Hydrolases; C067959:rhamnogalacturonase A