Abstract:

According to the PEPT1 gene sequence in GenBank,specific primer sequences were designed to amplify the PEPT1 epitope fragment gene.The PEPT1 epitope gene was subcloned into expression vector pET-22b(+)to give pET-22b-PEPT1 and expressed in Rosetta(DE3).The expressed product was purified by Ni-NTA chromatography column and identified by SDS-PAGE,Western blot and mass spectrogram.This study was beneficial for further research on producing specific antibody for poultry intestinal PEPT1.

Key words: chicken, PEPT1 gene, Rosetta（DE3）, expression vector;

摘要：

根据GenBank的原鸡PEPT1基因保守区序列设计引物,PCR扩增得到PEPT1基因抗原表位区序列,将该序列定向克隆至pET22b(+)载体上,成功构建了pET-22b-PEPT1重组质粒。重组质粒在原核表达宿主Rosetta(DE3)中诱导表达出PEPT1抗原表位区蛋白,纯化的表达产物经过质谱分析鉴定,为进一步生产鸡肽转运载体PEPT1的特异性抗体奠定了基础。

关键词: 鸡 PEPT1基因 Rosetta（DE3） 表达载体