Molecular cloning and sequencing of the gene encoding signal peptide for antibody of swine and construction of its eucaryotic expression vector

A fragment about 85 bp was amplified by RT_PCR technique from the genome mRNA of lymphocyte from swine,using primers designed and synthesized according to the gene sequence encoding signal peptide for antibody of swine.The amplifed fragments were cloned into pGEM_T vector and then sequenced.The determination of nucleotide sequence showed that the gene encoding signal peptide for antibody of swine spanned 57 bp,encoding 19 amino acids and the nucleotide and amino acid sequence of the gene cloned was the same as the reported sequences,and in 3'end understream of the gene encoding signal peptide for antibody of swine the signal peptidase cleavage site and cloning sites for foreign gene could be provided by the recombinant plasmid.and then the gene encoding signal peptide for antibody of swine was subcloned into pcDNA3.1(+) and verified by PCR and restriction endonuclease digestion,the recombinant eucaryotic expression vector of the p3.1_SFc was successfully constructed.This result laid foundation for further study of the recombinant eucaryotic expression vector to produce and secrete a foreign protein from living cells and study of the function of gene.