弓形虫膜表面抗原SAG1基因的原核表达及重组蛋白的免疫诊断价值  

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

作　　者：王朝兰[1] 汤冬生[1] 姚湧[2] 汪学龙[2] 王业梅[1] 

机构地区：[1]安徽中医学院寄生虫学教研室,合肥230038 [2]安徽医科大学病原生物学教研室

出　　处：《中国地方病学杂志》2011年第4期376-378,共3页Chinese Jouranl of Endemiology

摘　　要：目的 探讨重组弓形虫膜表面抗原SAG1基因的表达产物-原核表达蛋白（rSAG1）用于弓形虫病的免疫诊断价值.方法 用异丙基-B-D-硫代吡喃半乳糖苷（IPTG）诱导大肠埃希菌重组质粒pET28a-SAG1（pET28a-SAG1/BL21）表达,纯化重组pET28a-SAG1/BL21弓形虫膜表面抗原SAG1基因表达产物;用弓形虫缓殖子感染的鼠血清、正常鼠血清和10例弓形虫患者血清为一抗,基因表达产物rSAG1用免疫印迹法进行鉴定,比较rSAG1在弓形虫病免疫诊断中的价值.结果 纯化重组弓形虫膜表面抗原SAG1基因后获得了相对分子质量约38.5×103的表达产物rSAG1;表面抗原SAG1可被弓形虫缓殖子感染的鼠血清所识别;10例弓形虫患者血清在免疫印迹诊断中,有4例出现了弓形虫膜表面抗原SAG1基因表达产物rSAG1.结论 rSAG1具备一定的弓形虫病的免疫诊断价值.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.

关 键 词：弓形虫 膜表面抗原 重组蛋白 免疫诊断 

分 类 号：R686[医药卫生—骨科学]