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目的制备弓形虫微线体蛋白8羧基端胞质尾(MIC8t)蛋白片段。方法以弓形虫基因组为模板,PCR扩增MIC8t基因片段,构建MIC8t/pGEX-4T-1原核表达系统;IPTG诱导表达GST-MIC8t融合蛋白;亲和层析纯化融合蛋白。结果构建了MIC8t原核表达系统,表达并纯化了GST-MIC8t融合蛋白。结论在体外制备并纯化了GST-MIC8t融合蛋白,为后续研究MIC8t的功能奠定了基础。
Objective To prepare Toxoplasma gondii protein 8 carboxyterminal tail (MIC8t) protein fragment. Methods Toxoplasma gondii genome was used as a template to amplify the MIC8t gene fragment by PCR. The prokaryotic expression system of MIC8t / pGEX-4T-1 was constructed. IPTG induced the expression of GST-MIC8t fusion protein and purified the fusion protein by affinity chromatography. Results The MIC8t prokaryotic expression system was constructed and the GST-MIC8t fusion protein was expressed and purified. Conclusion The GST-MIC8t fusion protein was prepared and purified in vitro, which laid the foundation for the further study on the function of MIC8t.