沙打旺黄矮根腐病菌钙调素基因的克隆  

Cloning of calmodulin gene from Embellisia astragali

作　　者：刘建利[1,2] 李彦忠[1,3] 

机构地区：[1]草地农业生态系统国家重点实验室兰州大学草地农业科技学院,甘肃兰州730020 [2]北方民族大学生物科学与工程学院,宁夏银川750021 [3]中国农业科学院草原研究所,内蒙古呼和浩特010010

出　　处：《草业科学》2015年第5期719-724,共6页Pratacultural Science

基　　金：中国农业科学院科技创新工程(CAAS-ASTIP-IGR-2015-03);国家自然科学基金(31272496);长江学者和创新团队发展计划(PCSIRT);公益性行业(农业)科研专项经费项目(201303057)

摘　　要：沙打旺埃里砖格孢(Embellisia astragali)引起的黄矮根腐病是造成沙打旺(Astragalus adsurgens)草地退化最严重的病害之一。钙调素是Ca2+介导的信号通路中重要的分子。本研究旨在从沙打旺埃里砖格孢中克隆钙调素编码基因序列。以基因组DNA为模板,采用简并引物CAL-228F和CAL-737R扩增得539bp的同源片段;采用Hi-TAIL PCR分别获得344bp的5′侧翼序列和729bp的3′侧翼序列,拼接获得基因的1 290bp的全长DNA序列;利用全长引物CaMF和CaMR,得到840bp的DNA全长序列和450bp的cDNA全长序列,DNA全长序列中有4个内含子,均具有典型的AT-AG特征,cDNA全长序列翻译出149氨基酸多肽。The yellow stunt and root rot of standing milk-vetch disease caused by Embellisia astragali is one of the major factors in the pasture degradation.Calmodulin is significant protein in Ca2+dependent signaling pathway.The present study was aimed to clone calmodulin gene fromE.astragali genomic DNA.The homologous DNA fragment of 539 bp was amplified from genomic DNA with primers CAL-228 Fand CAL-737 R.Then,the 5′flanking sequence of 344 bp and 3′flanking sequence of 729 bp were amplified using Hi-TAIL PCR.Finally,the 1 290 bp full-length DNA sequence was assembled and 840 bp fulllength cDNA sequence was amplified with primers CaMF and CaMR.There were 4introns in DNA fulllength sequence which had the typical characteristics of AT-AG.The polypeptide with 149 amino acid was translated from cDNA sequence.

关 键 词：Hi-TAIL PCR 沙打旺埃里砖格孢 钙调素 基因克隆 

分 类 号：S435.4[农业科学—农业昆虫与害虫防治]