风疹病毒E1蛋白原核可溶性表达、纯化及血清学诊断研究

Soluble Expression and Purification in Prokaryotic System and Research on Serological Diagnosis of Rubella Virus E1 Protein

目的通过基因工程的方法获得风疹病毒E1蛋白在原核系统内可溶性表达，亲和纯化后，评价该重组蛋白在风疹病毒患者血清学诊断方面的可行性。方法将PCR扩增得到的风疹病毒E1核酸序列克隆至原核表达载体pET-NusA中，利用原核系统表达融合蛋白NUSA-E1，用WesternBlot和ELISA方法对其抗原性进行初步评价。结果NUSA-E1蛋白以可溶性形式进行表达，亲和纯化后可以获得高纯度的重组蛋白。WesternBlot和EI。ISA方法结果均证明该重组蛋白具有较强的抗原活性。结论融合蛋白NusA-E1在大肠埃希菌中以可溶性形式表达，高纯度的重组融合蛋白NUSA—E1抗原性强，临床上可以作为血清学诊断抗原，检测风疹病毒抗体。

Objective To obtain Rubella Virus E1 protein antigen expression in solubility in prokaryotic system through gene engineering method. After affinity purification,its feasibility for serological diagnosis was evaluated. Methods E1 gene was obtained by PCR and cloned into pET-NusA fusion vector,fusion protein NusA-E1 got expressed in prokaryotic system. The antigenicity of antigen of the product were examined by Western Blot and ELISA assay. Results NusA-E1 fusion protein was expressed in solubility in E. coli and high purity recombinant protein was obtained by affinity furification. The results of Western Blot and ELISA showed that the recombinant protein had strong antigenic ability. Conclusion The results suggest that the NusA-E1 fusion protein could expresse in solubility in Escherichia coll. The strong antigenieity of recombinant protein may be a potential candidate to be a serodiagnostic reagent to detect RV antibody.