禽致病性大肠杆菌pilA基因的克隆及其在乳酸菌中的表达

Cloning and expression of avian pathogenic Escherichia coli pilA gene in Lactococcus lactis

根据GenBank中禽致病性大肠杆菌pilA基因序列设计合成1对引物,以本实验室分离的禽致病性大肠杆菌基因组DNA为模板,采用PCR技术扩增得到pilA基因片段,经测序鉴定准确后将其克隆到乳酸乳球菌表达载体pMG36e中,构建重组质粒并将其电转入乳酸乳球菌MG1363,得到重组乳酸乳球菌。SDS-PAGE分析显示,表达的蛋白约为19 ku,与预期相符。Western blot进一步证实了该蛋白的免疫反应性。

To clone and express the pilA gene of avian pathogenic Escherichia coli, a pair of primers were designed according to the pilA gene sequences in GenBank. The PCR product was cloned into pMG36e vector. The recombinant expression plasmid was constructed. Then the recombinants were transformed into the host strain MG1363. The SDS-PAGE analysis showed that a 19ku protein was expressed. Western blot further confirmed its immunoreactivity.