摘要
目的构建人重组ICOS-Linker-Ig真核表达载体并预测其二级结构及Linker的合理性。方法扩增人I-COS胞外域及IgG Fc段,设计一段柔性Linker将二者连接。经测序分析得到融合基因序列。应用核酸和蛋白质序列分析软件EditSeqTM对融合基因及Linker部位翻译后二级结构水平的一些生物学特性,诸如柔性、抗原性、亲水性及表位等加以预测分析。结果ICOS-Linker-Ig融合基因的氨基酸序列经软件分析,并分别与ICOS和IgG单独分析结果比较,发现在二级结构水平未出现新的抗原性,同时Linker部位具有很低的抗原性,亲水性无改变,Linker部位呈中性且柔性良好,不影响两端蛋白质二级结构和融合蛋白空间构象。ICOS和Ig具有各自原来的表位特征,无新表位出现。结论本研究构建的ICOS-Linker-Ig能够保留ICOS和Ig各自生物活性和功能,计算机分析表明翻译后不影响蛋白质空间结构的形成,适合融合蛋白功能的发挥。
Objective To construct the expressed recombinant ICOS-Linker-Ig and predict the rationality and feasibility of the linker. Methods Human ICOS and IgG Fc fragment were produced by polymerase chain reaction and were combined by a flexible Linker. Utilize the sequence analysis software to analyze the flexibility, antigenicity, hoop and woods hydrophilicity and episode of recombinant ICOS-Linker-Ig. Results After the analysis by the software, it could be found that the recombinant gene has correct domains of ICOS and Ig. The linker had low episode, low antigenicity and high flexibility. Conclusion The results of computer analysis could help to rationally design the recombinant ICOS-Linker-Ig and keep its maximum biological activity.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2005年第6期479-482,共4页
Journal of Harbin Medical University
基金
哈尔滨医科大学研究生创新基金资助(2005)
