Centis S, Dumas B, Fournier J, Marolda M, Esquerré-Tugayé M T
Université Paul Sabatier, Centre de Biologie et Physiologie Végétale URA 1941 CNRS, Toulouse, France.
Gene. 1996 Apr 17;170(1):125-9. doi: 10.1016/0378-1119(95)00867-5.
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.
根据炭疽菌（Cl）β小种内切多聚半乳糖醛酸酶（EndoPG）的N端氨基酸（aa）序列以及不同真菌EndoPG中保守的内部序列设计的寡脱氧核糖核苷酸引物，用于聚合酶链反应（PCR）以扩增该真菌的基因组相关序列。获得了一个542 bp的片段，命名为pgA，并用作探针筛选Cl的部分基因组文库。在阳性克隆中，对其中一个进行了进一步分析。该克隆的核苷酸测序揭示了一个开放阅读框（ORF），其编码一个363个氨基酸（aa）的多肽，起始于一个26 aa的信号肽，该信号肽被一个70 bp的内含子打断，并且与十个真菌EndoPG序列具有高度同源性。在5'非编码区鉴定出了共有序列。此后，该基因组克隆被命名为Clpg1。用Clpg1特异性探针进行的Southern分析表明，该基因在Cl基因组中以单拷贝形式存在。
