论文部分内容阅读
目的克隆弓形虫RH株致密颗粒蛋白2(GRA2)的全长基因,在大肠埃希菌工程菌中重组表达GST-HisGRA2融合蛋白。方法提取弓形虫总RNA,反转录获得cDNA,PCR扩增GRA2基因全长,与pET-41Ek/LIC载体通过基因重组直接连接,在大肠埃希菌RosettaTM(DE3)pLysS工程菌中诱导表达重组融合蛋白,然后采用SDS-PAGE和Western blot鉴定重组蛋白产物。结果 PCR鉴定弓形虫GRA2全基因重组表达质粒构建正确;诱导表达的GSTHis-GRA2融合重组蛋白分子质量单位为52ku,与预测值相符。结论本研究成功构建了弓形虫GRA2全基因重组蛋白质粒,该质粒能表达GRA2蛋白,为研究GRA2与宿主细胞的相互作用奠定了基础。
OBJECTIVE: To clone the full-length gene of GRA2 from RH strain Toxoplasma gondii, and express GST-HisGRA2 fusion protein in Escherichia coli engineered bacteria. Methods The total RNA of Toxoplasma gondii was extracted and the cDNA was obtained by reverse transcription. The full length of GRA2 gene was amplified by PCR and ligated with pET-41Ek / LIC vector through gene recombination to induce expression and recombination in Escherichia coli RosettaTM (DE3) pLysS Fusion protein, and then using SDS-PAGE and Western blot identification of recombinant protein products. Results PCR identification of Toxoplasma gondii GRA2 whole gene recombinant plasmid was constructed correctly. The molecular mass unit of GSTHis-GRA2 fusion recombinant protein induced by PCR was 52ku, which was consistent with the predicted value. Conclusions The recombinant plasmids of GRA2 gene of Toxoplasma gondii were successfully constructed in this study. GRA2 protein can express GRA2 protein, which lays the foundation for studying the interaction between GRA2 and host cells.