刚地弓形虫表面膜抗原SAG1基因的原核表达及重组蛋白的诊断价值

Q3 Medicine

中华地方病学杂志 Pub Date : 2011-07-20 DOI:10.3760/CMA.J.ISSN.1000-4955.2011.04.007

Chao-lan Wang, D. Tang, Yong Yao, Xue-long Wang, Yean Wang

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引用次数: 0

摘要

目的探讨重组表面抗原1 (rSAG1)在弓形虫病免疫诊断中的价值。方法采用异丙基β-D- 1 -硫代半乳糖苷(IPTG)诱导大肠埃希菌重组质粒pET28a-SAG1 (pET28a-SAG1/BL21)的表达。pET28a-SAG1/BL21的表达产物(rSAG1)经Western blotting鉴定。以刚地弓形虫速殖子感染小鼠血清、正常小鼠血清和10例刚地弓形虫患者血清作为抗刚地弓形虫初级抗体，采用Western blotting方法鉴定rSAG1基因产物，比较rSAG1在弓形虫病中的诊断价值。结果经诱导纯化得到rSAG1蛋白，其相对分子质量为38.5 × 103。该融合蛋白可被感染弓形虫速殖子的小鼠血清识别，10例患者中4例检测到弓形虫表面膜抗原SAG1基因表达产物rSAG1。结论rSAG1在弓形虫病的免疫诊断中具有潜在的应用价值。关键词:刚地弓形虫;表面抗原;重组蛋白;免疫诊断

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Prokaryotic expression of surface membrane antigen SAG1 gene from Toxoplasma Gondii and the diagnostic value of the recombinant protein

Objective To investigate the diagnostic value of the recombinant surface antigen 1 (rSAG1) in immunodiagnosis of toxoplasmosis. Methods Isopropyl β-D- 1 -thio-galaetopyranoside (IPTG) was used to induce the expression of recombinant plasmid pET28a-SAG1 of Escherich coli(pET28a-SAG1/BL21 ). The expression products (rSAG1) of pET28a-SAG1/BL21 were identified by Western blotting. The serum of mice infected with Toxoplasma gondii tachyzoites, normal mouse serum and the serum from 10 toxoplasma gondii patients were used as primary anti-Toxoplasma gondii antibodies, and the rSAG1 gene products were identified by Western blotting, by which the diagnostic value of rSAG1 in Toxoplasmosis was compared. Results After induction and purification, rSAG1 protein was obtained and its relative molecular mass was 38.5 × 103. The fusion protein could be recognized by the serum of mouse infected with Toxoplasma gondii tachyzoites, rSAG1 of expression products of surface membrane antigen SAG1 gene from Toxoplasma Gondii could be detected in 4 cases from 10 patients by Westem blotting.Conclusion The rSAG1 has a potential value in the immunodiagnosis of Toxoplasmosis. Key words: Toxoplasma gondii; Surface antigen; Recombinant protein; Immunodiagnosis

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来源期刊

中华地方病学杂志我国对人类健康危害特别严重的地方性疾病：克山病、大骨节病、碘缺乏病、地方性氟中毒、地方性砷中毒、鼠疫、布鲁氏菌病、寄生虫、新冠肺炎等疾病，同时还报道多发性自然疫源性疾病。

CiteScore

1.60

自引率

0.00%

发文量

8714

期刊介绍： The Chinese Journal of Endemiology covers predominantly endemic diseases threatening health of the people in the areas affected by the diseases including Keshan disease, Kaschin-Beck Disease, iodine deficiency disorders, endemic fluorosis, endemic arsenism, plague, epidemic hemorrhagic fever, brucellosis, parasite diseases and the diseases related to local natural and socioeconomic conditions; and reports researches in the basic science, etiology, epidemiology, clinical practice, control as well as multidisciplinary studies on the diseases.