南方农业学报2018，Vol.49Issue(2)：360-366,7.DOI:10.3969/j.issn.2095-1191.2018.02.24
广西巴马小型猪SP1基因克隆测序及其真核表达载体的构建
Clone sequencing of gene SP1 in Guangxi Bama mini pig and construction of its eukaryotic expression vector
摘要
Abstract
[Objective]The purpose of this study was to clone specificity protein 1(SP1)gene of Guangxi Bama mini pig,construct itseukaryotic expression vector and lay a foundation for revealing its regulation mechanism to pig growth.[Method]Gene SP1 was amplified from longissimus dorsi of Guangxi Bama mini pig by RT-PCR.Bioinformatics analysis on the gene was conducted using DNASTAR and TMHMM.Eukaryotic expression vector pEGFP-N1-SP1 was construct-ed by plasmid pEGFP-N1 and was transfected into 293T cell.After 48 h of transfection,it was observed and taken pic-tures under inverted fluorescence microscope.[Result]The cloned Guangxi Bama mini pig SP1 gene sequence was consis-tent with wild boar reference sequence(XM_005652569.3)in GenBank(Sus scrofa),and no base mutation site was found.The coding sequence(CDS)was 2340 bp in length,coding 779 amino acids.SP1 protein of Guangxi Bama mini pig contained no transmembrane structure and signal peptide,and was not secretory protein.In its secomdary structure, there were 17.84% α-helix,21.31% extension strand,9.76% β-turn and 51.09% random coil.Green fluorescence protein has been found in 293T cells with transfected eukaryotic expression vector pEGFP-N1-SP1.[Conclusion]The encoded protein of gene SP1 CDS sequence in Guangxi Bama mini pig is mainly involved in intracellular activity,but it is not exo-crine protein.The eukaryotic expression vector pEGFP-N1-SP1 which is constructed based on gene SP1 is transfected into 293T cells and successfully expressed in it. It can be used in further research in function and interference expression of gene SP1.关键词
广西巴马小型猪/SP1基因/克隆/真核表达载体/生物学信息分析Key words
Guangxi Bama mini pig/gene SP1/clone/eukaryotic expression vector/bioinformatics analysis分类
农业科技引用本文复制引用
张广杰,崔悦悦,邱庆庆,夏琴,李龙,司景磊,夏攀杰,邹辉,兰干球..广西巴马小型猪SP1基因克隆测序及其真核表达载体的构建[J].南方农业学报,2018,49(2):360-366,7.基金项目
国家现代农业产业技术体系广西生猪产业创新团队建设项目(nycytxgxcxtd-03-15) （nycytxgxcxtd-03-15）
