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本研究利用3’-RACE技术从百脉根中克隆了一个钙调素基因(Lj Ca M4),并利用荧光定量PCR技术对该基因在根、茎、叶、花和豆荚中的相对表达量进行了分析。结果显示,扩增得到642 bp的片段,开放阅读框为453 bp,3’非翻译区(3’-UTR)长189 bp,编码150个氨基酸,分子量约为17.13 k D,理论等电点为4.03。多序列比对显示Lj Ca M4与其它植物中的Ca Ms有较高的保守性,与蒺藜苜蓿的同源性高达91%。Lj Ca M4m RNA在百脉根各组织中均有分布,其中在根中表达量最高,依次为根>茎>叶>豆荚>花。本研究成功克隆并鉴定了百脉根Lj Ca M4基因,初步检测了该基因在百脉根各组织器官中的表达差异,为进一步探明该基因可能的生物学功能提供了一定的理论依据。
In this study, a calmodulin gene (Lj Ca M4) was cloned from Lotus japonicus by 3’-RACE technique. The relative expression level of this gene in roots, stems, leaves, flowers and pods Analyzed. The results showed that a 642 bp fragment was obtained. The open reading frame was 453 bp. The 3 ’untranslated region (3’-UTR) was 189 bp in length and encoded a protein of 150 amino acids with a molecular mass of 17.13 kD. The theoretical isoelectric point 4.03. Multiple sequence alignment showed that Lj Ca M4 was highly conserved among other plants, with 91% homology with Medicago truncatula. Lj Ca M4m RNA was distributed in all tissues of Lotus corniculatum, among which the highest expression was in root> stem> leaf> pod> flower. In this study, we successfully cloned and identified Lj Ca M4 gene from Lotus japonicus, and preliminarily detected its expression in tissues and organs of Lotus cornea. This study provides a theoretical basis for further exploration of the possible biological function of this gene.