番茄gdp - d -甘露糖焦磷酸化酶cDNA的克隆、表达及定位

ZOU Li-Ping , LI Han-Xia , OUYANG Bo , ZHANG Jun-Hong , YE Zhi-Biao
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引用次数: 10

摘要

gdp - d -甘露糖焦磷酸化酶(GMP, EC 2.7.7.22)催化gdp - d -甘露糖的合成,是植物抗坏血酸生物合成的第一步。以马铃薯GMP cDNA序列为查询探针,从GenBank的dbEST中获得65条高度同源的番茄ESTs序列,并对推定的番茄GMP cDNA序列进行了组装。利用RACE-PCR技术克隆了番茄GMP全长cDNA,并根据组装后的cDNA序列设计引物。全长cDNA序列包含1 086 bp的完整开放阅读框(ORF),编码361个氨基酸残基。该基因被指定为LeGMP (GenBank登录号)。AY605668)。同源性分析表明,该氨基酸与马铃薯GMP同源性为96%,与马铃薯、烟草、苜蓿和拟南芥的GMP同源性分别为99%、97%、91%和89%。Northern blot分析表明,LeGMP在番茄根、茎、叶、花和果实中均有组成性表达;但表达水平各不相同。利用75个番茄渗入系(il)绘制了LeGMP的3-D图谱,每条渗入系包含一个纯合的rflp定义的染色体片段,这些染色体片段来自绿果物种Lycopersicon pennellii。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Cloning, Expression, and Mapping of GDP-D-mannose Pyrophosphorylase cDNA from Tomato (Lycopersicon esculentum)

GDP-D-mannose pyrophosphorylase (GMP, EC 2.7.7.22) catalyzes the synthesis of GDP-D-mannose and represents the first committed step in plant ascorbic acid biosynthesis. Using potato GMP cDNA sequence as a querying probe, 65 highly homologous tomato ESTs were obtained from dbEST of GenBank and the putative cDNA sequence of tomato GMP was assembled. The full-length GMP cDNA of tomato was cloned by RACE-PCR with primers designed according to the assembled cDNA sequence. The full-length cDNA sequence contained a complete open reading frame (ORF) of 1 086 bp, which encoded 361 amino acid residues. This gene was designated as LeGMP (GenBank accession No. AY605668). Homology analysis of LeGMP showed a 96% identity with potato GMP and the deduced amino acid showed 99%, 97%, 91% and 89% homology with GMP from potato, tobacco, alfalfa and Arabidopsis thaliana, respectively. Northern blot analysis showed that LeGMP was constitutively expressed in roots, stems, leaves, flowers and fruits of tomato; but the expression levels varied.LeGMP was mapped to 3-D using 75 tomato introgression lines (ILs), each containing a single homozygous RFLP-defined chromosome segment from the green-fruited species Lycopersicon pennellii.

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