Construction of Double-Gene Expression Vector and Its Biological Effects

The purpose is to promote proliferation of intestinal epithelial cells,accelerate the rehabilitation of intestinal mechanical barrier and prevent intestinal bacteria translocation.We cloned the proglucagon cDNA including GLP-2 gene sequence by polymerase chain reaction and reverse transcription.The signal peptides and mature peptides of GLP2 which were produced by PCR were linked by SOE(splicing by overlap extension)The second amino acid in the N-terminal of GLP-2,was mutated from GCT(coded alanine)to GGT(coded glycin)by PCR.The human gene defensin 5 was cloned from pcDNA3.1-HD-5-cDNA,then we constructed the GLP-2 and HD-5 cDNA into double-gene expression vector pVITRO3 and tranfected intestinal epithelium cells to observe the ability of proliferation and anti-bacterium of IEC-6 and Caco-2 cells.The results showed that construction of double gene expression vector had the ability of prompting proliferation of cells and inhibiting the multiplication and adherence of bacteria.