Isolation and sequence analysis of Clpg1, a gene coding for an endopolygalacturonase of the phytopathogenic fungus Colletotrichum lindemuthianum.
Oligodeoxyribonucleotide primers designed from the N-terminal amino acid (aa) sequence of the endopolygalacturonase (EndoPG) of Colletotrichum lindemuthianum (Cl) race beta and from an internal sequence conserved among different fungal EndoPG were used in a polymerase chain reaction (PCR) to amplify genomic related sequences of the fungus. A 542-bp fragment, designated pgA, was obtained and used as a probe to screen a partial genomic library of Cl. Among the positive clones, one was further analyzed. Nucleotide sequencing of this clone revealed on ORF encoding a 363-amino-acid (aa) polypeptide beginning with a signal peptide of 26 aa interrupted by an intron of 70 bp, and showing a high degree of homology to ten fungal EndoPG sequences. Consensus sequences were identified in the 5' non-coding region. This genomic clone was thereafter designated Clpg1. Southern analysis, performed with a Clpg1-specific probe, showed that this gene is present as a single copy in the Cl genome.
Centis
S
S
Université Paul Sabatier, Centre de Biologie et Physiologie Végétale URA 1941 CNRS, Toulouse, France.
Dumas
B
B
Fournier
J
J
Marolda
M
M
Esquerré-Tugayé
M T
MT
D001483:Base Sequence; D004271:DNA, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics*; D015723:Gene Library; D005800:Genes, Fungal; D003904:Mitosporic Fungi / Q000235:genetics*; D008969:Molecular Sequence Data; D011096:Polygalacturonase / Q000235:genetics*; D021382:Protein Sorting Signals / Q000235:genetics; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D004271:DNA, Fungal; D005656:Fungal Proteins; D021382:Protein Sorting Signals; D011096:Polygalacturonase