Reymond P, Deléage G, Rascle C, Fèvre M
Laboratoire de Biologie Cellulaire Fongique, Université Lyon I, CGMC-CNRS UMR 106, Villeurbanne, France.
Gene. 1994 Sep 2;146(2):233-7. doi: 10.1016/0378-1119(94)90298-4.
The phytopathogenic fungus Sclerotinia sclerotiorum produces a number of extra-cellular pectin-degrading enzymes. We have cloned and determined the complete sequence of a gene (pg1) encoding an endopolygalacturonase (PG1). The coding region consists of a non-interrupted 1143-bp open reading frame. S. sclerotiorum pg1 was compared to other fungal PG-encoding genes. Basic transcription control sequences were identified in the 5' non-coding region. The deduced amino acid (aa) sequence (380 aa) of the enzyme is compared to seven fungal PG sequences and shows a high level of identity (41.5 to 59.8%). Predicted secondary structures were compared, revealing a similar protein organization most probably in antiparallel beta sheets. Hybridization analysis using a pg1 0.65-kb BamHI fragment as a probe allowed the identification of seven different recombinant phages from a genomic library. Analysis of the hybridizing restriction fragments suggests that PG-encoding genes are organized as a family.
植物致病真菌核盘菌能产生多种胞外果胶降解酶。我们已克隆并测定了一个编码内切多聚半乳糖醛酸酶（PG1）的基因（pg1）的完整序列。编码区由一个不间断的1143碱基对的开放阅读框组成。将核盘菌的pg1与其他真菌的PG编码基因进行了比较。在5′非编码区鉴定出了基本转录控制序列。该酶推导的氨基酸序列（380个氨基酸）与7个真菌PG序列进行了比较，显示出高度的同源性（41.5%至59.8%）。对预测的二级结构进行了比较，揭示了最可能以反平行β折叠形式存在的相似蛋白质结构。使用pg1的0.65kb BamHI片段作为探针进行杂交分析，从基因组文库中鉴定出7种不同的重组噬菌体。对杂交限制性片段的分析表明，PG编码基因是作为一个家族组织的。
