摘要
目的:克隆、表达结核分枝杆菌PPE家族蛋白PPE68的编码基因Rv3873,并初步探索PPE68诱导Balb/c小鼠脾淋巴细胞的增殖活性,为进一步研究PPE68的免疫功能提供实验依据。方法:以结核分枝杆菌H37Rv基因组为模板,PCR扩增Rv3873基因编码序列,定向克隆入融合蛋白原核表达载体pET32a(+),获得重组表达质粒,转化大肠杆菌后用IPTG进行诱导表达,纯化表达产物,通过XTT比色法检测重组蛋白PPE68诱导脾淋巴细胞的增殖活性。结果:重组质粒pETRv3873构建成功,57kDa的His-PPE68融合蛋白在大肠杆菌中稳定表达,并具有刺激Balb/c小鼠脾淋巴细胞增殖的功能。结论:PPE68蛋白具有较强的免疫原性,其免疫学功能具有进一步研究的价值。
Objective, To express and purify recombinant PPE68, and study the proliferation activity of splenic lymphocyte induced by recombinant PPE68. Methods： Rv3873 was amplified from Mycobacterium tuberculosis H37Rv genomic DNA by PCR. PCR product was cloned into prokaryotic expression vector pET32a（ ＋ ）. The recombinant plasmid with PPE68 gene was transformed into E. coli BL21 （DE3） to express the fusion protein His-PPE68 induced by IPTG . Stimulating lymphocyte of spleen with purified recombinant PPE68 and detecting lymphocyte proliferation activity by XTT colorimetry rnethod. Results： Recombinant expression vector pETRv3873 was successfully constructed and the recombinant plasmid could express the 57 kDa fusion protein His-PPFA58 stably. Conclusion： It has the potentM of inducing splenic lymphocyte proliferation, providing the basis for the further study of immunogenicity and immunoprotection of PPE68 from TB infection.
出处
《四川生理科学杂志》
2005年第2期49-51,共3页
Sichuan Journal of Physiological Sciences
基金
国家自然科学基金资助(30271172)
作者简介
王晓琳(1976-)，女。辽宁铁岭人。硕士。主要从事微生物分子生物学与免疫学功能研究;通讯作者：鲍朗(E-mail：baolan@weurns．Edu．Cn)
