人EPO-Linker-EGFP融合蛋白的设计和生物活性预测

The design of hEPO-Linker-EGFP fusion protein and its biology characteristics prediction

目的 探讨hEPO -Linker -EGFP融合蛋白设计的合理性。方法 应用GeneConstructionKit2 5 ,www .expasy .com网站提供的分析方案 ,分析重组体的开放读框及融合蛋白的柔性 ,并作了蛋白质二级结构模拟。结果 重组体的转录受四环素应答顺式作用元件调控 ,Linker所在部位柔性高 ,融合蛋白表达后 ,二级结构预测Linker不改变蛋白结构 ,完全符合作者设计hEPO -Linker-EGFP融合蛋白的初衷。结论 重组蛋白设计合理 ,融合蛋白有很大可能保留了hEPO和EGFP的理化特性 ,为进一步的实践操作提供了可靠的理论基础。

Objective To study the reasonability of designation for hEPO-Linker-EGFP fusion protein. Methods Using sequence analysis software and protocols prescribed on website www. expasy. com to analyze the open reading frame of the recombinant and the flexibility, as well as the secondary structure of hEPO-linker-EGFP fusion protein. Results The transcription of the recombinant protein was regulated by the tetracycline-responsed cisacting element. The fusion protein had correct domains of hEPO and EGFP. Linker had high flexibility and did not influence the secondary structure of fusion protein. Conclusion Computer analysis can help to rationally design the recombinant protein. The attenuated hEPO and EGFP protein keep their maximum biological activities.