SARS病毒核衣壳蛋白的表达及其疫苗的免疫原性研究

发布时间：2018-03-25 19:05

本文选题：SARS　切入点：核衣壳蛋白　出处：《吉林大学》2005年硕士论文

【摘要】：本研究人工合成了编码SARS 冠状病毒(SARS-CoV)核衣壳蛋白的基因序列(1296bp),并构建了分别在大肠杆菌、毕赤酵母、哺乳动物细胞中表达的重组质粒pET-28a-N、pPIC9K-N、pIRES-N。证明上述重组质粒在大肠杆菌、毕赤酵母中均能表达SARS-CoV 核衣壳蛋白,另进行了该重组蛋白的表达条件优化和纯化。将核酸疫苗质粒pIRES-N 转染至BHK 细胞证明其表达SARS-CoV N 蛋白后,将该重组质粒免疫小鼠,证明能够诱导抗SARS-CoV N 蛋白抗体,流式细胞仪检测表明,鼠脾T 淋巴细胞数量增多,具有较好的细胞免疫诱导作用。将大量制备的上述核酸疫苗注射马体,免疫三次,并应用大肠杆菌、毕赤酵母中表达的核衣壳蛋白进行了加强免疫。证明了重组核酸疫苗免疫后,可诱导较高水平的抗SARS-CoV N 蛋白的抗体,另应用细菌和酵母中表达的N 蛋白追加免疫可诱导更高水平的抗SARS-CoV N 蛋白抗体,这为大量制备治疗用抗SARS 抗体奠定了良好基础。
[Abstract]:In this study, the nucleocapsid protein encoding SARS coronavirus (SARS-CoV) was synthesized and cloned into E. coli, Pichia pastoris and mammalian cells respectively. SARS-CoV nucleocapsid protein was expressed in Pichia pastoris and the expression conditions of the recombinant protein were optimized and purified. After transfection of nucleic acid vaccine plasmid pIRES-N into BHK cells to prove its expression of SARS-CoV N protein, mice were immunized with the recombinant plasmid, which was proved to be able to induce anti-#en3# N protein antibody. Flow cytometry analysis showed that the number of spleen T lymphocytes increased. It has good cellular immune induction effect. The nucleic acid vaccine was injected into the horse body three times, and the nucleocapsid protein expressed in Escherichia coli and Pichia pastoris was used to enhance the immunization. The antibody against SARS-CoV N protein could be induced at a higher level, and a higher level of anti SARS-CoV N protein antibody could be induced by supplementary immunization with N protein expressed in bacteria and yeast, which laid a good foundation for the preparation of anti SARS antibody for treatment.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392.1;Q78

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