Hohnjec N, Becker J D, Pühler A, Perlick A M, Küster H
Biologie VI (Genetik), Universität Bielefeld, Germany.
Mol Gen Genet. 1999 Apr;261(3):514-22. doi: 10.1007/s004380050995.
We have isolated and sequenced a sucrose synthase (SucS) cDNA from the model legume Medicago truncatula. This cDNA (MtSucS1) contains an ORF of 2418 bp, coding for a protein of 805 amino acids with a molecular mass of 92.29 kDa. The deduced amino acid sequence shows significant homology to other plant sucrose synthases, in particular to the nodule-enhanced sucrose synthases from pea and broad bean. Northern analysis revealed that the corresponding gene shows a ten-fold higher expression level in root nodules than in uninfected root, stem and leaf tissues. SucS protein was detected in root nodules from a variety of legumes, including M. truncatula. Whereas only one SucS isoform was detectable in root nodules, an additional sucrose synthase of slightly larger molecular weight was present in uninfected root, stem and flower tissues of M. truncatula. From our expression and sequence data we infer that the MtSucS1 gene encodes a nodule-enhanced sucrose synthase in M. truncatula. Southern hybridization data indicate that MtSucS1 is a single-copy gene. An analysis of a genomic MtSucS1 sequence revealed that the gene consists of 14 exons with the start codon being located on exon II. As is common for SucS genes, the MtSucS1 gene contains a large intron of 747 bp in the 5' untranslated region. The transcriptional start of MtSucS1 was mapped and putative regulatory elements in the MtSucS1 promoter were identified.
我们从模式豆科植物蒺藜苜蓿中分离并测序了一个蔗糖合酶(SucS)cDNA。这个cDNA(MtSucS1)包含一个2418 bp的开放阅读框,编码一个由805个氨基酸组成、分子量为92.29 kDa的蛋白质。推导的氨基酸序列与其他植物蔗糖合酶具有显著同源性,特别是与豌豆和蚕豆的根瘤增强型蔗糖合酶。Northern分析表明,相应基因在根瘤中的表达水平比未感染的根、茎和叶组织高10倍。在包括蒺藜苜蓿在内的多种豆科植物的根瘤中检测到了SucS蛋白。虽然在根瘤中只能检测到一种SucS同工型,但在蒺藜苜蓿未感染的根、茎和花组织中存在另一种分子量稍大的蔗糖合酶。根据我们的表达和序列数据,我们推断MtSucS1基因在蒺藜苜蓿中编码一种根瘤增强型蔗糖合酶。Southern杂交数据表明MtSucS1是一个单拷贝基因。对基因组MtSucS1序列的分析表明,该基因由14个外显子组成,起始密码子位于外显子II上。与SucS基因常见情况一样,MtSucS1基因在5'非翻译区含有一个747 bp的大内含子。确定了MtSucS1的转录起始位点,并鉴定了MtSucS1启动子中的推定调控元件。