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水稻(Oryza sativa L.)磷酸葡萄糖异构酶编码cDNA的特性分析。

Characterization of cDNA encoding for phosphoglucose isomerase of rice (Oryza sativa L.).

作者信息

Nozue F, Umeda M, Nagamura Y, Minobe Y, Uchimiya H

机构信息

Institute of Molecular and Cellular Biosciences, University of Tokyo, Japan.

出版信息

DNA Seq. 1996;6(3):127-35. doi: 10.3109/10425179609010200.

Abstract

Two types of genes (Pgi-a and Pgi-b) encoding phosphoglucose isomerase (PGI; EC 5.3.1.9) were cloned from cDNA libraries of rice cultured cells (Oryza sativa L.). Pgi-a and Pgi-b consisted of 2132 and 2030 nucleotides, respectively. The homology between these genes was 93.0% at nucleotide level. The homology scores between these genes in protein coding region and 3' non-coding region were 95.6% and 79.4%, respectively. PGI proteins encoded by Pgi-a and Pgi-b consisted of 567 and 568 amino acid, respectively, sharing 95.8% homology at amino acid sequences. Of 11 PGI genes from other plant species and organisms whose amino acid sequences had been determined, a dicotyledonous plant Clarkia lewisii PGI showed the highest homology (about 80%) with rice PGIs. GC contents at the third position of rice PGI genes were about 40%. In order to confirm the enzyme activity of the protein encoded by the rice cDNA, Pgi-a was subcloned into an expression vector, pBluescript II SKp, which was introduced into Escherichia coli. The transformant had an additional PGI activity from Pgi-a.

摘要

从水稻培养细胞(Oryza sativa L.)的cDNA文库中克隆出了两种编码磷酸葡萄糖异构酶(PGI;EC 5.3.1.9)的基因(Pgi-a和Pgi-b)。Pgi-a和Pgi-b分别由2132和2030个核苷酸组成。这些基因在核苷酸水平上的同源性为93.0%。这些基因在蛋白质编码区和3'非编码区的同源性得分分别为95.6%和79.4%。由Pgi-a和Pgi-b编码的PGI蛋白分别由567和568个氨基酸组成,在氨基酸序列上具有95.8%的同源性。在已确定氨基酸序列的来自其他植物物种和生物体的11个PGI基因中,双子叶植物Clarkia lewisii PGI与水稻PGI的同源性最高(约80%)。水稻PGI基因第三位的GC含量约为40%。为了确认水稻cDNA编码蛋白的酶活性,将Pgi-a亚克隆到表达载体pBluescript II SKp中,并导入大肠杆菌。该转化体具有来自Pgi-a的额外PGI活性。

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