百脉根钙调素基因LjCaM4的3'-RACE克隆及其表达量分析  

Cloning by 3'-RACE and Expression of LjCaM4 Gene in Lotus japonicus

作　　者：舒茂荣 秦宗志 陈杨晗 汤加勇[2] 廖进秋[1] 

机构地区：[1]四川农业大学生命科学学院,雅安625014 [2]四川农业大学动物营养研究所,温江611130

出　　处：《基因组学与应用生物学》2017年第8期3057-3064,共8页Genomics and Applied Biology

基　　金：四川省教育部重点项目(No.14ZA0008)资助

摘　　要：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 the research, a calmodulin gene （LjCaM4） was cloned by 3＇-RACE from Lotus japonicus. The expression of it in root, stem, leaf, flower and pod were analyzed using quantitative real-time PCR. The results showed that LjCaM4 including 189 bp 3＇-teriminal UTR and 453 bp open reading frame （OR_F）, which is 642 bp and encodes 150 amino acids. Its theoretical molecular weight is 17.13 kD, and its isoelectric point is 4.03. Multiple sequence alignment indicates that these CaMs are highly conservation among species and LjCaM4 has 91% sequence identity with that of Medicago truncatula. Furthermore, real-time PCR analyses demonstrate that the expression of LjCaM4 in roots is significantly higher than that in the other tissues, and the relative expression levels are root 〉stem 〉leaf〉bean-pod〉flower. In the research, LjCoM4 was successfully cloned and identified in L. japonicas. Its relative expression in different tissues were detected by qRT-PCR. These could provide theoretical basis for the future study in biological function of LjCaM4.

关 键 词：百脉根 LJ CA M4 3'-RACE 荧光定量PCR 

分 类 号：Q943.2[生物学—植物学] S541.6[农业科学—作物学]