盐生草Actin基因片段的克隆及表达  

Cloning and expression analysis of Actingene fragment from halophyte Halogeton glomeratus

作　　者：马艳红[1] 徐先良[1] 汪军成[1] 任盼荣 杨柯[1,2] 孟亚雄[1,2] 李葆春[1,3] 马小乐[1,2] 王化俊[1,2] 

机构地区：[1]甘肃省干旱生境作物学重点实验室/甘肃省作物遗传改良与种质创新重点实验室,甘肃兰州730070 [2]甘肃农业大学农学院,甘肃兰州730070 [3]甘肃农业大学生命科学技术学院,甘肃兰州730070

出　　处：《草业科学》2015年第9期1432-1437,共6页Pratacultural Science

基　　金：973计划前期研究专项(2014CB160313);国家自然科学基金面上项目(31171558);国家自然科学基金地区项目(31460347);国家大麦青稞产业技术体系(CARS-05);甘肃省自然基金(1208RJZA136)

摘　　要：根据藜科植物的肌动蛋白(Actin)基因编码区保守序列设计一对简并引物,提取盐生草(Halogeton glomeratus)叶片的总RNA,运用RT-PCR技术扩增出Actin基因的核心序列并连接到pMD19-T载体,阳性克隆经PCR检测后进行测序。序列分析表明,盐生草Actin基因核心序列长度为598bp,编码198个氨基酸,并在GenBank中注册,登录号为KF699314,由盐生草Actin基因推导的氨基酸序列与其他几种耐盐植物的同源性均在94%以上,具有高度的保守性。采用荧光定量RT-PCR技术分析其Actin基因在盐生草不同器官的表达情况,结果显示其表达量恒定无差异,表明克隆的Actin基因可作为盐生草内参基因来研究其相关耐盐基因的表达。In the present study, the Actin gene fragment was amplified by reverse transcription polymerase chain reaction （RT-PCR） from total RNA extracted from leaves of H. glomeratus with designed degenera- ted primers based on the conserved sequences of Actin genes from chenopodiaceous plants. The Actin gene fragments were cloned into pMD19-T vector, sequenced using positive clone and registered in GenBank （accession number：KF699314）. The sequence results indicated that the length of Actin gene fragment was 598 bp which encoded 198 amino acids and homology of amino acid sequences translated from nucleotides acids from H. glomeratus and other plants was 94% which suggested the high conservation of this gene. There was no difference between Actin gene expressions in different organs of H.glomeratus based on quantitative RT-PCR （qRT-PCR） analysis which suggested that Actin gene can be selected as internal con- trol gene for study the expression of other salt-tolerant genes of H. glomeratus.

关 键 词：盐生草 肌动蛋白基因 克隆 序列分析 

分 类 号：Q943.2[生物学—植物学]