稻瘟病菌Dam1基因的克隆、原核表达及纯化  

Cloning, Prokaryotic Expression and Purification of Dam1 Gene from Magnaporthe oryzae

作　　者：许雨晨 吕哲[1] 陈保善[1,2] 彭好文[1,2] Xu Yuchen Lv Zhe Chen Baoshan Peng Haowen(College of Agriculture, Guangxi University, Nanning, 530001 State Key Laboratory for Conservation and Utilization of Subtropical Agro-biore- sources, Guangxi University, Nanning, 530001)

机构地区：[1]广西大学农学院,南宁530001 [2]广西大学亚热带农业生物资源保护与利用国家重点实验室,南宁530001

出　　处：《基因组学与应用生物学》2016年第9期2385-2389,共5页Genomics and Applied Biology

基　　金：国家自然科学基金(31160356);广西自然科学基(2014GXNSFAA118101)共同资助

摘　　要：稻瘟病菌(Magnaporthe oryzae(Hebert)Barr)是一种植物病原真菌,可引发水稻的毁灭性病害—稻瘟病,在全世界范围内造成了极大的经济损失。寻找有效防治稻瘟病的方法迫在眉睫。本课题组在前期研究中发现了一种新的稻瘟病菌致病相关基因——Mo Dam1。本研究以稻瘟病菌的RNA反转录得到的c DNA为模板克隆Dam1的ORF,并将其和p ET-30a连接,构建原核表达载体p ET-30a-Dam1,之后转化到大肠杆菌BL21(DE3)中,用IPTG诱导表达Dam1重组蛋白,最终纯化后得到Dam1蛋白。Rice blast fungus Magnaporthe oryzae is a kind of plant pathogenic fungus. It can cause rice blast, which is one of the catastrophic disease of rice and make a great economic loss worldwide. Now it is imminent to find effective methods for contolling the rice blast. We found a new kind of rice blast fungus pathogenesis related genes in prior studies--Daml. The ORF of Dam1 gene was amplified by recerse transcription-polymerase chain reaction from total RNA extracted from Magnaporthe oryzae. Through the construction of pET-3Oa-Daml vector, the recombinant Daml protein was successfully expressed by IPTG in Escherichia coli BL21 （DE3） cells.

关 键 词：稻瘟病菌 稻瘟病 Dam1 原核表达 蛋白纯化 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]