论文部分内容阅读
弓形虫棒状体蛋白 1(ROP1)是与虫体侵入相关的重要分子。将含编码 ROP1蛋白基因的真核表达重组质粒 pc DNA3转染 Hep G- 2细胞 ,为进一步研究做准备。用脂质体介导的真核细胞转染法 ,G418筛选阳性克隆 ,SDS-PAGE及 Western- blot分析鉴定表达产物。结果显示 ,在所筛选的克隆化生长的细胞裂解液样品电泳图谱中 ,有一大小约 35～ 40 k Da的特异电泳条带 ,该条带可被弓形虫免疫兔血清识别。本研究建立了体外稳定表达 ROP1的细胞系 ,证实了表达产物的免疫原性
Toxoplasma rod protein 1 (ROP1) is an important molecule involved in invasion of parasites. The recombinant plasmid pcDNA3 containing the gene encoding ROP1 gene was transfected into Hep G-2 cells for further study. The positive clones were screened by G418 with liposome-mediated eukaryotic cell transfection, and the expressed products were identified by SDS-PAGE and Western-blot analysis. The results showed that there was a specific electrophoresis band about 35 ~ 40 kDa in the cloned cell lysis solution of the screened samples, which was recognized by Toxoplasma gondii immunized rabbit serum. This study established a cell line stably expressing ROP1 in vitro and confirmed the immunogenicity of the expressed product