Xu Y, Hall T C
Institute of Developmental and Molecular Biology, Texas A&M University, College Station 77843-3155.
Plant Physiol. 1993 Feb;101(2):683-7. doi: 10.1104/pp.101.2.683.
A cDNA clone encoding rice (Oryza sativa L.) cytosolic triosephosphate isomerase (TPI), an important glycolytic enzyme, was isolated and characterized. The clone (pRTPI-6) contains an open reading frame of 759 base pairs, encoding a polypeptide chain of 253 amino acid residues (M(r) 27,060). The identity of this clone was defined by its high homology (85% nucleotide sequence and 89% amino acid sequence identical match) with a maize mRNA sequence encoding the cytosolic TPI and with TPIs from other species. Genomic DNA blot analysis using the cDNA as a probe showed that the cytosolic TPI gene is present as a single copy per haploid rice genome, as opposed to that found in maize, in which multiple TPI gene copies exist. A single TPI mRNA species of about 1100 nucleotides was detected by gel blot hybridization analysis of RNA isolated from root, culm, and leaf tissues, indicating that its expression is ubiquitous. Based on sequence comparison and molecular analysis, we propose that the chloroplast-located TPI may be encoded by divergent structural nuclear genes in rice.
一个编码水稻(Oryza sativa L.)胞质磷酸丙糖异构酶(TPI)的cDNA克隆被分离和鉴定,该酶是一种重要的糖酵解酶。该克隆(pRTPI - 6)包含一个759个碱基对的开放阅读框,编码一条由253个氨基酸残基组成的多肽链(M(r) 27,060)。通过与编码胞质TPI的玉米mRNA序列以及其他物种的TPI具有高度同源性(核苷酸序列85%相同匹配,氨基酸序列89%相同匹配)来确定该克隆的身份。以该cDNA为探针进行的基因组DNA印迹分析表明,与玉米中存在多个TPI基因拷贝不同,胞质TPI基因在每个单倍体水稻基因组中以单拷贝形式存在。通过对从根、茎和叶组织中分离的RNA进行凝胶印迹杂交分析,检测到一种约1100个核苷酸的单一TPI mRNA种类,表明其表达具有普遍性。基于序列比较和分子分析,我们提出水稻中位于叶绿体的TPI可能由不同的结构核基因编码。