本文選題：SARS　切入點：核衣殼蛋白　出處：《吉林大學》2005年碩士論文

【摘要】：本研究人工合成了編碼SARS 冠狀病毒(SARS-CoV)核衣殼蛋白的基因序列(1296bp),并構建了分別在大腸桿菌、畢赤酵母、哺乳動物細胞中表達的重組質粒pET-28a-N、pPIC9K-N、pIRES-N。證明上述重組質粒在大腸桿菌、畢赤酵母中均能表達SARS-CoV 核衣殼蛋白,另進行了該重組蛋白的表達條件優(yōu)化和純化。 將核酸疫苗質粒pIRES-N 轉染至BHK 細胞證明其表達SARS-CoV N 蛋白后,將該重組質粒免疫小鼠,證明能夠誘導抗SARS-CoV N 蛋白抗體,流式細胞儀檢測表明,鼠脾T 淋巴細胞數(shù)量增多,具有較好的細胞免疫誘導作用。 將大量制備的上述核酸疫苗注射馬體,免疫三次,并應用大腸桿菌、畢赤酵母中表達的核衣殼蛋白進行了加強免疫。證明了重組核酸疫苗免疫后,可誘導較高水平的抗SARS-CoV N 蛋白的抗體,另應用細菌和酵母中表達的N 蛋白追加免疫可誘導更高水平的抗SARS-CoV N 蛋白抗體,這為大量制備治療用抗SARS 抗體奠定了良好基礎。
[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.
【學位授予單位】：吉林大學
【學位級別】：碩士
【學位授予年份】：2005
【分類號】：R392.1;Q78

【參考文獻】

相關期刊論文 前2條

1 ;The Structure Analysis and Antigenicity Study of the N Protein of SARS-CoV[J];Genomics Proteomics & Bioinformatics;2003年02期

2 ;Identification of an epitope of SARS-coronavirus nucleocapsid protein[J];Cell Research;2003年03期

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本文編號：1664448