摘要
目的 :在大肠杆菌中表达变形链球菌葡聚糖结合蛋白B基因 (glucanbindingproteinB ,gbpB) ,并对重组蛋白进行初步纯化 ,为下一步制备多抗和进行相关的研究奠定基础。方法 :将克隆到的gbpB全长编码区基因克隆到表达载体 pPROEXTMHTb ,构建表达质粒 pPROEXTMHTb - gbpB ,以大肠杆菌E .coliBL2 1(DE3)为宿主菌进行诱导表达 ,表达产物经镍 -次氮基三乙酸 (Ni-NTA)柱进行亲和层析纯化。结果 :工程菌经IPTG诱导 3h后 ,在SDS -PAGE上出现一条新的蛋白条带 ,相对分子量 (Mr)为 4 .5～ 5 .0× 10 3 ,以可溶形式存在 ,经Ni-NTA柱纯化后获得纯度较高的重组葡聚糖结合蛋白B。结论 :获得Mr为 4 .5～ 5 .0× 10 3
Objective: To express and purify the recombinant glucan binding protein B of Streptococcus mutans in Escherichia coli. Methods: The DNA fragment encoding the glucan binding protein B of S. mutans was inserted into expression vector pPROEX TMHTb following ATG and a 6 histidine for affinity purification. The recombinant plasmid was transformed into E. coli BL21 (DE3) and induced by IPTG to express fusion protein. The recombinant protein was purified by metal-chelate affinity chromatography on Ni-NTA resin. Results: After induction with IPTG for 3 hour, a new foreign protein band near Mr 4.5～5.0×10 3 appeared on SDS-PAGE. Conclusion: The recombinant glucan binding protein B of S. mutans was successfully expressed in E. coli BL21 (DE3) and purified. These make it possible for further study.
出处
《口腔医学研究》
CAS
CSCD
2004年第4期340-342,共3页
Journal of Oral Science Research
关键词
变形链球菌
葡聚糖结合蛋白B
基因表达
蛋白纯化
Streptococcus mutans Glucan binding protein B Gene espression Protein purification
