摘要
目的制备小鼠模型疫苗HBV S-ecdCD40L融合基因表达载体并采用软件预测其所表达蛋白的合理性及生物活性。方法首先从质粒中扩增HBV S基因,并从小鼠淋巴细胞中扩增CD40L胞外段(extracellular domain,ECD)基因。然后通过RT-PCR法将其串联并与载体相连接,完成构建后采用软件分析其所表达蛋白是否与预期一致。结果构建小鼠模型表达载体并通过软件分析HBV S、ecdCD40L两个基因连接之后其蛋白的疏水性和亲水性无改变,也没有出现新的表位。连接链(linker)不影响融合蛋白的二级结构。结论该载体的设计符合预期,其表达的融合蛋白保留了HBV S和小鼠CD40L胞外段的生物学活性,为进一步研究奠定了基础。
Objective To construct murine HBV S - ecdCD40L expression vector by biotechnology and forecast the biological activi- ty of HBV S - ecdCD40L fusion protein. Methods First HBV S gene was amplified from plasmid and mouse ecdCD40L from lympho- eytes, then HBV S genes was fused and murine eedCD40L genes in RT -PCR and expression vector was constructed. Whether the ex- pressed proteins in line with expectations was analyzed by software. Results The expression vector was constructed successfully. The hy- drophobic and hydrophilic of fusion protein was the same and fusion protein secondary structure level had no new epitope. The linker did not influence the secondary structure of the protein. Conclusion The design of the fusion protein is appropriate. It keeps the biological activities of HBV S and murine ecdCD40L.
出处
《医学研究杂志》
2013年第12期67-70,共4页
Journal of Medical Research
基金
温州市科技局基金资助项目(Y20100204)
