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目的为分析多头带绦虫Tm18重组蛋白的免疫原性。方法利用RT-PCR技术从激活的多头带绦虫六钩蚴中扩增Tm18基因,将扩增产物亚克隆入pGEX-4T-1载体中,构建pGEX-4T-rTm18表达载体,转化至大肠杆菌BL21中,经IPTG诱导表达融合蛋白;表达产物进行SDS-PAGE和Western-blot分析后,将纯化的Tm18重组蛋白免疫小鼠,用ELISA检测其血清抗体。结果多头带绦虫Tm18基因序列长为552 bp,包括1个396 bp的开放性阅读框。表达的重组蛋白大小约为39.4 kDa,与脑包虫患羊血清的免疫印迹分析呈阳性反应;小鼠免疫1周后血清中即可检测到抗Tm18蛋白的特异性抗体,并于免疫后第5周达到高峰。结论 Tm18重组蛋白具有较好的免疫原性,可作为脑包虫疫苗的候选抗原。
Objective To analyze the immunogenicity of Tm18 recombinant protein of tapeworm. Methods The Tm18 gene was amplified by RT-PCR from Acinetobacter baumannii. The amplified product was subcloned into pGEX-4T-1 vector to construct pGEX-4T-rTm18 expression vector and transformed into E. coli BL21 The fusion protein was induced by IPTG. After the expressed product was analyzed by SDS-PAGE and Western-blot, the purified Tm18 recombinant protein was immunized with the antibody and the serum antibody was detected by ELISA. Results The length of Tm18 gene was 552 bp in length, including a 396 bp open reading frame. The expressed recombinant protein was about 39.4 kDa in size, which was positive with Western blot analysis of the brain echinococcosis sheep serum. Anti-Tm18 protein-specific antibody was detected in serum one week after immunization, 5 weeks to reach the peak. Conclusion Tm18 recombinant protein has good immunogenicity and can be used as a candidate antigen for brain hydatid vaccine.