Isolation and sequence analysis of Clpg1, a gene coding for an endopolygalacturonase of the phytopathogenic fungus Colletotrichum lindemuthianum
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- 作者：Centis ; Sylvie ; Dumas ; Bernard ; Fournier ; Joë ; lle ; Marolda ; Michè ; le ; Esquerré ; -Tugayé ; Marie-Thé ; rè ; se
- 关键词：Pectin degradation ; pectinase ; polymerase chain reaction ; nucleotide sequence ; sequence comparison ; Southern blot analysis
- 刊名：Gene
- 出版年：1996
- 出版时间：April 17, 1996
- 年：1996
- 卷：170
- 期：1
- 页码：125-129
- 全文大小：517 K
文摘
Oligodeoxyribonucleotide primers designed from the N-terminal amino acid (aa) sequence of the endopolygalacturonase (EndoPG) of Colletotrichum lindemuthianum (Cl) race β 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 an ORF encoding a 363-amino-acid (aa) polypeptide begining 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.