论文部分内容阅读
目的应用基因工程方法构建表达幽门螺杆菌尿素酶Ｂ亚单位（ＵｒｅＢ）的菌株．方法用ＰＣＲ方法从幽门螺杆菌染色体ＤＮＡ上扩增出ＵｒｅＢ基因片段，将其克隆至ｐＳＫ（＋）质粒上，然后插入原核表达载体ｐＥＴ２２ｂ（＋）中，在大肠杆菌ＢＬ２１（ＤＥ３）中表达．结果经测序ＵｒｅＢ片段有１７０７ｂｐ组成，为编码５６９个氨基酸残基的多肽．ＳＤＳＰＡＧＥ和免疫印迹分析检测发现，ＵｒｅＢ基因表达的蛋白质分子质量为６５０００Ｕ，并证实该重组蛋白质可以被幽门螺杆菌感染阳性患者的血清所识别，同时将其免疫小鼠可刺激机体产生抗该重组蛋白质的抗体．结论重组的ＵｒｅＢ可以作为一种有效的蛋白质疫苗用于幽门螺杆菌感染的预防和治疗．
Objective To construct a strain expressing UreB of Helicobacter pylori by genetic engineering. Methods The UreB gene fragment was amplified from Helicobacter pylori by PCR and cloned into pSK (+) plasmid. The recombinant plasmid was inserted into prokaryotic expression vector pET22b (+) and expressed in Escherichia coli BL21 (DE3 ). Results The sequenced UreB fragment was composed of 1707bp and encoded a polypeptide of 569 amino acid residues. SDS-PAGE and Western Blot analysis showed that the molecular weight of UreB gene expressed protein was 65000U, and confirmed that the recombinant protein can be recognized by the serum of H. pylori-positive patients, while immunized mice can stimulate the body to produce anti- Recombinant protein antibodies. Conclusion Recombinant UreB can be used as an effective protein vaccine in the prevention and treatment of Helicobacter pylori infection.