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来自植物病原真菌围小丛壳菌的甘油醛-3-磷酸脱氢酶编码基因及cDNA的克隆与分子特征分析

Cloning and molecular characterization of the glyceraldehyde-3-phosphate dehydrogenase-encoding gene and cDNA from the plant pathogenic fungus Glomerella cingulata.

作者信息

Templeton M D, Rikkerink E H, Solon S L, Crowhurst R N

机构信息

Molecular Genetics Group, Horticulture and Food Research Institute, New Zealand Ltd., Mt. Albert, Auckland.

出版信息

Gene. 1992 Dec 1;122(1):225-30. doi: 10.1016/0378-1119(92)90055-t.

DOI:10.1016/0378-1119(92)90055-t
PMID:1452034
Abstract

The glyceraldehyde-3-phosphate dehydrogenase gene (gpdA) has been identified from a genomic DNA library prepared from the plant pathogenic fungus Glomerella cingulata. Nucleotide sequence data revealed that this gene codes for a putative 338-amino-acid protein encoded by two exons of 129 and 885 bp, separated by an intron 216 bp long. The 5' leader sequence is also spliced by an intron of 156 bp. A cDNA clone was prepared using the polymerase chain reaction, the sequence of which was used to confirm the presence of the intron in the coding sequence and the splicing of the 5' leader sequence. The transcriptional start point (tsp) was mapped at -253 nt from the site of the initiation of translation by primer extension and is adjacent to a 42-bp pyrimidine-rich region. The general structure of the 5' flanking region shows similarities to gpdA from Aspergillus nidulans. The putative protein product is 71-86% identical at the aa level to GPDs from Aspergillus nidulans, Cryphonectria parasitica, Curvularia lunata, Podospora anserina and Ustilago maydis.

摘要

已从植物致病真菌炭疽菌(Glomerella cingulata)制备的基因组DNA文库中鉴定出3-磷酸甘油醛脱氢酶基因(gpdA)。核苷酸序列数据显示,该基因编码一种推定的338个氨基酸的蛋白质,由129 bp和885 bp的两个外显子编码,中间间隔一个216 bp长的内含子。5'前导序列也由一个156 bp的内含子进行剪接。使用聚合酶链反应制备了一个cDNA克隆,其序列用于确认编码序列中内含子的存在以及5'前导序列的剪接。通过引物延伸将转录起始点(tsp)定位在距翻译起始位点-253 nt处,且与一个42 bp富含嘧啶的区域相邻。5'侧翼区域的总体结构与构巢曲霉(Aspergillus nidulans)的gpdA相似。推定的蛋白质产物在氨基酸水平上与来自构巢曲霉、栗疫霉(Cryphonectria parasitica)、新月弯孢霉(Curvularia lunata)、黑麦草核腔菌(Podospora anserina)和玉米黑粉菌(Ustilago maydis)的GPD有71%-86%的同一性。

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