大菱鲆hepcidin抗菌肽基因的克隆及在大肠杆菌中的高效表达  

Cloning and High-level Expression of A Turbot Hepcidin Antimicrobial Peptide Gene

作　　者：李伟[1] 陈松林[2] 赵晓杰[2] 

机构地区：[1]长江大学生命科学学院,湖北荆州434025 [2]农业部海洋渔业资源可持续利用重点开放实验室中国水产科学研究院黄海水产研究所,山东青岛266003

出　　处：《长江大学学报（自科版）（中旬）》2007年第3期84-87,共4页Journal of Yangtze University(Nature Science Edition)

基　　金：国家自然科学基金资助项目(40376047)

摘　　要：通过同源克隆结合RACE的方法从大菱鲆(Scophthal mus maximus)肝脏中克隆了大菱鲆hepcidin抗菌肽基因全长cDNA(GenBank accession number AY994074)。大菱鲆hepcidin基因cDNA全长778bp,包含有1个273bp的开放阅读框,编码1个长90氨基酸的前体肽。将大菱鲆hepcidin抗菌肽基因的成熟肽序列重组至原核表达载体pGEX-4T-1中,用IPTG诱导后,SDS-PAGE电泳显示在29ku处有特异性蛋白条带出现,表达的融合蛋白主要以包涵体形式存在,融合蛋白约占总蛋白的26%。Western-blotting分析发现表达的融合蛋白可以特异性地与GST抗体相识别。这些结果表明大菱鲆hepcidin抗菌肽基因成功实现了原核表达。The molecular cloning of a hepcidin antimicrobial peptide gene from the liver of turbot using RACE were studied.The full-length cDNA of turbot hepcidin is 778 bases long and contains an ORF of 273 bases encoding a prepropeptide of 90 amino acid residues.The hepcidin mature peptide sequence was amplified and inserted into pGEX-4T-1 and fusion expression plasmid was constructed.The plasmid was transferred into E.coli BL21(DE3)plyS and induced by IPTG.SDS-PAGE and western-blotting analysis showed that the recombinant had expressed a 29 ku specific protein successfully.

关 键 词：大菱鲆(Scophthalmus maximus) HEPCIDIN 抗菌肽 表达 

分 类 号：Q785[生物学—分子生物学]