| 摘要: |
| 扩增了炭疽芽孢杆菌PA基因，构建了原核表达质粒pET-28a-PA，并对其进行了诱导表达，对表达产物免疫原性和免疫保护效果进行了研究。结果表明，PA基因的长度约为2 205 bp，PA蛋白的相对分子质量为83 ku；rPA蛋白具有良好的免疫原性，初免后10 d即可检测到特异性抗体，第3次免疫后抗体水平明显升高，可达1∶128 00；rPA蛋白刺激小鼠产生的特异性抗体亚型以IgG1为主；rPA蛋白在体外对小鼠脾脏细胞有促进生长和增殖作用。rPA免疫组小鼠脾脏中CD+4细胞显著增加，免疫应答趋向Th2型反应，说明rPA介导的免疫反应以体液免疫为主。rPA具有一定的免疫保护作用，保护率达到50％。 |
| 关键词: 炭疽芽孢杆菌 保护性抗原 免疫 基因工程疫苗 |
| DOI： |
| 分类号: |
| 基金项目:国家“973”计划项目(2002CB513208) |
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| Cloning and immunoprotection of PA recombinant protein of Bacillus anthracis |
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| Abstract: |
| PA gene amplified was from Bacillus anthracis by PCR and then cloned into pET28a(+) vector.The recombinant plasmid pET28a(+)PA was constructed and expressed and the immunogenicity and immunoprotection were studied.The results show that the length of PA gene is about 2 205 bp and relative molecular mass of the expressed protein is 83 ku;the rPA has good immunogenicity,and specificity of antibody could be detected after 10 d immunization,and the antibody levels rise markedly,3rd to 1∶2 800;the rPA protein stimulates the specificity of antibody binding to mouse IgG1-based;the rPA protein can promote the growth and multiplication of the mouse spleen cells in vitro.The rPA immunization group can significantly increase CD+4 mouse spleen cells and the immune responses tend to Th2-type response,showing that the rPA most mediate to humoral immunity.The rPA is fairly immune protective,the protection rate being 50%. |
| Key words: Bacillus anthracis protective antigen immunity genetic engineering vaccine |
