紫花苜蓿叶酸合成关键酶GCHI和ADCS编码基因的克隆及分析

发布时间：2018-03-04 18:01

本文选题：紫花苜蓿　切入点：叶酸　出处：《植物生理学报》2017年07期 　论文类型：期刊论文

【摘要】：GTP环化水解酶I (GCHI)与氨基脱氧分支酸合酶(ADCS)是植物叶酸合成途径的限速酶,对叶酸的合成起着至关重要的作用。本研究利用cDNA末端快速扩增技术(RACE)结合逆转录PCR方法,克隆获得了紫花苜蓿(Medicago sativa)的GCHI和ADCS编码基因,分别命名为MsGCHI与MsADCS。序列分析结果表明,MsGCHI基因全长1 752 bp,包含1 371 bp的开放阅读框(ORF),编码456个氨基酸;MsADCS基因全长3 324 bp,ORF为2 613 bp,编码870个氨基酸。经同源比对与进化树分析,MsGCHI和Ms ADCS基因编码的氨基酸与蒺藜苜蓿(M.truncatula)的MtGCHI和MtADCS氨基酸序列具有最高的同源性,分别达到98%和96%,与其他植物的氨基酸序列相似度也均在58%以上。实时定量荧光PCR分析结果表明,MsGCHI与MsADCS基因在紫花苜蓿的根、茎、叶中均有表达,但在叶中的表达量都为最高。
[Abstract]:GTP cyclohydrolase I (I) and aminodeoxybranched acid synthase (ADCS) are rate-limiting enzymes for folic acid synthesis in plants, which play an important role in the synthesis of folic acid. In this study, rapid amplification of cDNA terminal was combined with reverse transcriptase (PCR). The GCHI and ADCS coding genes of Medicago sativa were cloned. They were named MsGCHI and MsADCS.The sequence analysis showed that the MsGCHI gene was 1 752 BP in length, containing 1 371 BP open reading frame (ORF), encoding 456 amino acids. The total length of 3 324 BP MsGCHI gene was 2 613 BP, encoding 870 amino acids. The amino acids encoded by MsGCHI and Ms ADCS genes were most homologous to the MtGCHI and MtADCS amino acid sequences of M. truncatula. The results of real-time quantitative PCR analysis showed that MsGCHI and MsADCS genes were expressed in the roots, stems and leaves of alfalfa, but they were the highest in leaves.
【作者单位】：兰州大学草地农业科技学院草地农业生态系统国家重点实验室;
【基金】：国家自然科学基金(31670405和31372360) 兰州大学中央高校基本科研业务费专项资金(lzujbky-2016-4)~~
【分类号】：Q943.2;S541.9

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