Thomas B R, Laudencia-Chingcuanco D, Gottlieb L D
Department of Genetics, University of California, Davis 95616.
Plant Mol Biol. 1992 Aug;19(5):745-57. doi: 10.1007/BF00027071.
The gene encoding a cytosolic isozyme of phosphoglucose isomerase (PGI, EC 5.3.1.9) was isolated from Clarkia lewisii, a wild flower native to California, and the structure and sequence of the entire coding region determined. PGI catalyzes an essential step in glycolysis and carbohydrate biosynthesis in plants. Spanning about 6 kb, the gene has 23 exons and 22 introns, the highest number yet reported in plants. The exons range in size from 43 to 156 nt and encode a protein of 569 amino acids. The protein is about 44-46% identical to the inferred protein sequences of pig, Escherichia coli and Saccharomyces cerevisiae. All of the introns are bordered with the consensus 5'-GT...AG-3' dinucleotides. The longest intron includes a large stem-loop structure bounded by a perfect 9 nt direct repeat. We cloned the PGI gene from a genomic library prepared from a single plant of known PGI genotype. The locus and allele of the clone were identified by matching restriction fragments to fragments from genetically defined genomic DNAs by Southern hybridization.
编码磷酸葡萄糖异构酶(PGI,EC 5.3.1.9)胞质同工酶的基因从原产于加利福尼亚的野生花卉刘易斯氏克拉克花中分离出来,并确定了整个编码区的结构和序列。PGI催化植物糖酵解和碳水化合物生物合成中的一个关键步骤。该基因跨度约6 kb,有23个外显子和22个内含子,是植物中迄今报道的数量最多的。外显子大小从43至156 nt不等,编码一个由569个氨基酸组成的蛋白质。该蛋白质与猪、大肠杆菌和酿酒酵母的推测蛋白质序列约有44%-46%的同源性。所有内含子都以保守的5'-GT...AG-3'二核苷酸为边界。最长的内含子包含一个由完美的9 nt直接重复序列界定的大茎环结构。我们从一株已知PGI基因型的单株植物制备的基因组文库中克隆了PGI基因。通过Southern杂交将限制性片段与来自遗传定义的基因组DNA的片段进行匹配,鉴定了克隆的基因座和等位基因。