Clone sequencing of gene SP1 in Guangxi Bama mini pig and construction of its eukaryotic expression vector

ObjectiveThe 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.MethodGene 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.ResultThe 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.ConclusionThe 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.