一个棉花黄萎病抗菌肽基因25a2的克隆与表达  

Cloning and Expression of a Cotton Verticillium Wilt Antibacterial Peptide Gene 25a2

作　　者：张冬冬[1] 王世英[1] 郭晓军[1] 姜军坡[1] 刘慧娟[1] 朱宝成[1] 

机构地区：[1]河北农业大学生命科学学院,保定071001

出　　处：《棉花学报》2009年第5期346-350,共5页Cotton Science

基　　金：河北省自然科学基金项目(398152)

摘　　要：将棉花黄萎病抗菌肽A2的N端15个氨基酸序列在NCBI进行同源序列比较,根据同源蛋白的DNA序列设计引物,以解淀粉芽孢杆菌(Bacillus amyloliquefaciens)RS-25菌株基因组为模板,应用PCR扩增出一条约300 bp的DNA片段。序列分析表明,DNA片段长354bp,编码117个氨基酸,将该基因命名为25a2。构建表达载体pET-28a-25a2,转化大肠杆菌(Eschrichia coli)BL21,37℃培养,应用终浓度为1 mmol.L-1的IPTG诱导,表达产物经SDS-PAGE分析,得到分子量约15 kDa表达蛋白条带,与理论蛋白分子量相符。In order to clone the gene which encodes the cotton Verticillium wilt antibacterial peptide A2,15 amino acid residues of N-terminal of the antibacterial peptide A2 was inputted in NCBI to blast and the result showed a protein contained a fragment which was the same with the 15 amino acid residues of A2.Following the protein sequence,the gene of the protein was found and a pair of primers was designed according to the gene.A DNA fragment about 300 bp was obtained by PCR amplification using the genome of stra...

关 键 词：棉花黄萎病 解淀粉芽孢杆菌 抗菌肽A2 

分 类 号：S435.621[农业科学—农业昆虫与害虫防治]