中文摘要：

利用基因工程的方法,以实验室保存的p MAL-C2X-NAP质粒为模板,PCR扩增NAP基因,构建重组质粒p GEX-NAP.重组质粒通过酶切和测序鉴定后转化大肠杆菌表达菌株BL21（DE3）,再经IPTG低温诱导获得可溶性GST-NAP融合蛋白,最后利用谷胱甘肽琼脂糖凝胶树脂进行纯化,Prescission蛋白酶进行柱上切割去除GST标签.结果表明,p GEX-NAP重组质粒构建正确,在大肠杆菌中经IPTG低温诱导表达,可获得大量可溶性GST-NAP融合蛋白.Prescission蛋白酶柱上切割去除GST标签后,经Western Blot验证NAP蛋白能被兔抗NAP多克隆抗体特异识别.

英文摘要：

To obtain plenty of HP-NAP（ H. pylori neutrophil-activating protein） for research,NAP gene was amplified from p MAL-C2X-NAP vector by PCR,and the recombinant plasmid p GEX-NAP was constructed. After verified by enzyme digestion and gene sequencing analysis,p GEX-NAP was transformed into E. coli BL21（ DE3） and the soluble fusion protein GST-NAP was obtained by inducing with IPTG in low temperature. Then GST-NAP was purified with Glutathione Sefinose Resin,and cleavaged GST-tag with Prescission protease on column. In conclusion,recombinant plasmid p GEX-NAP was successfully constructed and soluble GST-NAP fusion protein was efficiently expressed in E. coli. High purity HP-NAP can be obtained by Prescission protease cleavage on column. NAP protein was recognized by Rabbit antiNAP m Ab using Western Blot.