上海海洋大学学报2012，Vol.21Issue(4)：502-508,7.
草鱼抗病毒基因Mx全长cDNA的克隆、序列分析与真核表达载体构建
Cloning and sequence analysis of antiviral Mx gene full-length cDNA from grass carp （Ctenopharyngodon indellus） and its eukaryotic expression vector construction
摘要
Abstract
Mx protein is one of the classes of antiviral proteins induced by type I interferon （IFN）. A fulllength cDNA sequence of Mx gene from grass carp （Ctenopharyngodon indellus） was amplified using reverse transcription polymerase chain reaction （RT-PCR） with total RNA extracted from Ctenopharyngodon indellus kidney （CIK） cells treated With PolyI： C and cloned into pMD20-T vector named pMD20-T-Mx, and sequenced. The recombinant plasmid pMD20-T-Mx and eukaryotic expression vector pEGFP-C1 were cleaved by restriction endonuclease, Mx gene full-length cDNA sequence linkaged with pEGFP-C1. The data indicate that grass carp Mx gene full-length eDNA sequence has 1921 nucleotides, encoding 627 amino acid residues of the mature peptide with a molecular weight of about 71.1 ku and a theoretical isoelectric point of 8.33. The grass carp Mx protein contains a triparitite guanosine-5＇-Triphosphate （GTP）-binding motif and a dynamin family signature sequence, which are conserved in all IFN-indueed Mx proteins of the vertebrates. Bioinformatics analysis reveals that the protein encoded by Mx gene in grass carp is hydrophilic in most regions and has rich B cell antigenicity positions without obvious nuclear localization signal sequence. The dimensional structure of the grass carp Mx protein includes GTPase domain in the amino terminus region, central interactive domain （CID） in the middle and GTPase effector domain （GED） in the earboxyl terminus region. Comparative analysis shows that the Mx protein amino acid sequence of grass carp shares similarity of 45.1% - 99. 7% to those of other species, pEGFP-C1-Mx, which contains Mx gene full-length cDNA sequence and EGFP gene, has been constructed.关键词
草鱼/Mx基因/克隆/序列分析/载体构建Key words
grass carp/Mx gene/cloning/sequence analysis/vector construction分类
农业科技引用本文复制引用
刘莉,刘鹏,金佳丽,吴海丽,王改改,江晨,采克俊..草鱼抗病毒基因Mx全长cDNA的克隆、序列分析与真核表达载体构建[J].上海海洋大学学报,2012,21(4):502-508,7.基金项目
浙江省大学生科技创新活动计划（新苗人才计划）项目 （）
浙江省自然科学基金 （）
湖州市自然资金项目 （）
