Abstract: Objective Objective To clone and express the thioredoxin（Trx）from RH strain tachyzoites of Toxoplasma gondii，estab? lish the prokaryotic expression vector and purify the recombinant protein，then produce the polyclonal anti?Trx antibody in rab? bits. Methods Methods Trx fragment was amplified by PCR and cloned into the pET?28a （+） vector，and the recombinant protein was in? duced with IPTG and purified by Ni?NTA affinity chromatography. The polyclonal antibody specificity was detected by Western blotting. Results Results The trx gene was amplified from T. gondii cDNA by PCR. The recombinant plasmid trx/pET?28a （+）was use? fully constructed，and the recombinant TRX protein was expressed and purified. The TRX polyclonal antibody was also ob? tained. The specific band of TRX was detected by Western blotting. Conclusion Conclusion Western blotting can detect the specificity of polyclonal anti?Trx antibody，which will facilitate the biological functions of Trx.

Key words: Toxoplasma gondii, Thioredoxin, Cloning, Prokaryotic expression, Identification, Polyclonal antibody

摘要： 目的 目的 克隆刚地弓形虫硫氧还蛋白 （Thioredoxin，Trx） 基因， 构建原核表达载体， 通过诱导表达和纯化蛋白， 免 疫家兔制备多克隆抗体。方法 方法 采用PCR技术扩增刚地弓形虫Trx基因， 克隆至原核表达载体pET?28a （+） 中， 转化大肠 埃希菌 （E. coli） Rosetta， 用IPTG诱导目的蛋白表达， 采用镍亲和层析法获得纯化蛋白并免疫家兔制备多克隆抗体。利用 Western blotting技术鉴定多克隆抗体的特异性。结果 结果 成功从刚地弓形虫 cDNA 中扩增出 Trx 目的基因， 构建了 Trx/pET?28a （+） 重组质粒， 获得抗Trx重组蛋白的多克隆抗体。Western blotting技术检测出弓形虫Trx蛋白的特异性条 带。结论 结论 用制备的兔抗Trx多克隆抗体能检测弓形虫Trx在速殖子内的表达， 为进一步深入研究刚地弓形虫Trx功能 奠定了基础。

关键词: 刚地弓形虫, 硫氧还蛋白, 克隆, 原核表达, 鉴定, 多克隆抗体