期刊论文

中文

中华皮肤科杂志

0412-4030

2013

46

5

305-308

10.3760/cma.j.issn.0412-4030.2013.05.002

81201331:国家自然科学基金 11JJ4076:湖南省自然科学基金 11B107:湖南省教育厅青年项目 2011KJ32:衡阳市科技项目

本校为第一机构

目的探讨梅毒螺旋体重组蛋白TP0993在梅毒血清学诊断中的应用价值。方法通过生物信息学分析,获取TP0993基因序列,构建原核载体进行诱导表达;Ni-NTA亲合层析柱纯化重组蛋白,Western印迹检测其免疫抗原性,用重组蛋白直接注射免疫新西兰家兔,评价其免疫原性。用纯化的TP0993重组蛋白包被微孔板,建立间接酶联免疫吸附试验(ELISA)方法,检测临床梅毒患者血清和健康体检者正常血清,同时与梅毒螺旋体明胶凝集试验(TPPA)进行比较,根据重组蛋白与梅毒阴阳性血清的反应情况,评价重组抗原在梅毒血清学诊断中的应用价值。结果成功构建PET-28a(+)-0993原核表达载体,经表达、纯化后获得相对分子质量约为34 000的...

Objective To evaluate the value of a Treponema pallidum (TP) recombinant protein TP0993 in the serodiagnosis of syphilis.Methods A bioinformatics method was used to obtain the sequence of TP0993 gene.The open reading frame (ORF) without upstream non-coding region of TP0993 gene was ligated into the expression vector PET-28a (+),which was then transformed into Escherichia coli Rosetta.Isopropyl-β-d-thiogalactoside (IPTG) was used to induce the expression of TP0993 protein.The expressed protein was purified with nickel-nitrilotriacetic acid (Ni-...