蒙古冰草肌动蛋白基因片段的克隆与组织表达分析  

Cloning and analysis on tissue specific expression of a Actin gene fragment from Agropyron mongolicum

作　　者：云锦凤[1,2,3] 赵彦[1,2] 石凤敏[1] 王俊杰[1,2] 

机构地区：[1]内蒙古农业大学生态环境学院,内蒙古呼和浩特010018 [2]内蒙古自治区草品种育繁工程技术研究中心,内蒙古呼和浩特010018 [3]草地资源教育部重点实验室,内蒙古呼和浩特010018

出　　处：《草业学报》2011年第2期170-176,共7页Acta Prataculturae Sinica

基　　金：973项目子课题(2007CB108901-1);国家自然科学基金资助项目(30760159);内蒙古草业重大专项(20091401)资助

摘　　要：本研究利用同源序列法分离蒙古冰草Actin基因同源片段,并分析蒙古冰草Actin基因在根、茎、叶中的表达特征。根据禾本科植物小麦Actin基因的保守序列设计1对引物A1,经RT-PCR扩增,从蒙古冰草cDNA中克隆到1个长度为541 bp的Actin基因片段,使用DNAman和DNAUSER等分子生物学软件进行序列分析,结果表明,该序列编码179个氨基酸,并推测该片段具有Actin超基因家族的保守结构域,含有1个ATP结合位点,抑制蛋白结合位点和胶溶蛋白结合位点;该基因片段的氨基酸序列与小麦Actin基因的同源性为99%,与玉米同源性为96%,与水稻同源性为93%。组织器官特异性表达分析结果表明,该基因在根、茎、叶中的表达量恒定。将该基因片段命名为MwACT1,并在GengBank中登记注册,登录号为FJ490410。In this study,actin gene homology fragment from Mongo1ian wheatgrass（Agropyron mongolicum）was isolated using homology-based method,and analyzed on tissue specific expression of endogenous actin gene.The primers A1 was designed according to the amino acid conserved regions of the cloned actin gene of Gramineae plant wheat,one partials gene sequences have been obtained by reverse transcription polymerase chain reaction（RT-PCR）.Sequence analysis using Software of DNAman and DNAuser etc,and showed that the cDNA sequences was 541 bp,encoding 179 amino acids.And putative conserved domains have been detected,including ACTIN superfamily domains of ATP binding site,profilin binding site and gelsolin binding site.Homology comparison with Actin gene amino acid sequences in other plants showed that it was 99% identical to wheat,96% to maize and 93% to rice.The results of analysis on tissue specific expression showed that the endogenous actin gene expression level in the root,stem and leaf of Mongo1ian wheatgrass was identical.It was named as MwACT1,GenBank accession number： FJ490410.

关 键 词：蒙古冰草 ACTIN基因 克隆 表达分析 

分 类 号：S543.903[农业科学—作物学] Q943.2[生物学—植物学]