Construction and expression of three taste peptide tandem genes

According to the analysis of the structure of GD, AD and VD taste peptide, the tandem monomers were designed by introducing the specific sites for formic acid and trypsin.Oligonucleotides were designed and synthesized based on the code usage of Escherichia Coli and then 8-copy genes were structured by spliced, the corresponding genes were cloned into pET-30a which were finally transformed into BL21 (DE3) .The recombinants BL21-pET30-8GD, BL21-pET30-8AD, BL21-pET30-8VD were confirmed by positive clone screening and DNA sequencing.Expression of target genes was confirmed by modified Tricine-SDS-PAGE electrophoresis of engineering bacteria total protein and purified protein from supernatant of lysate.