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目的:构建金黄色葡萄球菌核酸酶(SN)基因的原核表达载体,研究其在大肠杆菌中的表达,并制备兔抗SN抗体。方法:从质粒pPLCSN中扩增SN基因片段,插入表达载体pLEX后转化大肠杆菌GI724,以色氨酸诱导表达。以表达的重组蛋白免疫日本大耳白兔制备抗SN抗体,用Westernblot分析兔抗SN抗体的特异性。结果:重组表达载体pLEXSN在大肠杆菌中获得表达,表达的可溶性蛋白占菌体蛋白总量的37%,具有良好的生物学活性。制备的兔抗SN抗体能够特异识别SN。结论:成功地在大肠杆菌中表达具有生物学活性的SN,并制备了兔抗SN抗体,为进一步探讨其作为抗病毒因子应用于病毒性疾病的治疗奠定了基础。
Objective: To construct a prokaryotic expression vector of Staphylococcus aureus nuclease (SN) gene, study its expression in E. coli and prepare rabbit anti-SN antibody. Methods: The SN gene fragment was amplified from the plasmid pPLCSN, inserted into the expression vector pLEX and transformed into E. coli GI724 to induce the expression of tryptophan. The anti-SN antibody was prepared by immunizing Japanese white rabbits with the expressed recombinant protein and the specificity of the rabbit anti-SN antibody was analyzed by Western blot. Results: The recombinant plasmid pLEXSN was expressed in Escherichia coli. The expressed soluble protein accounted for 37% of the total bacterial proteins and had good biological activity. The prepared rabbit anti-SN antibody can specifically recognize SN. CONCLUSIONS: The successful expression of SN with biological activity in E. coli and the preparation of rabbit anti-SN antibodies lay the foundation for the further study of its application as an antiviral agent in the treatment of viral diseases.