作　　者： ; ; ;

机构地区： 韶关学院英东生命科学学院

出　　处： 《南昌大学学报（理科版）》 2012年第6期552-555,共4页

摘　　要： 采用基因工程方法,将PhiX174噬菌体裂解基因E和温敏控制表达系统与质粒pPBA1100基因杂交,构建了重组子pPBA1100-E。将重组子转化到多杀性巴氏杆菌中,通过温度诱导使裂解基因表达。用限制性内切酶检验重组子,扫描电镜观察多杀性巴氏杆菌细菌幽灵和菌落形成单位评价遗传灭活率。结果表明:重组质粒经限制性内切酶酶切鉴定,琼脂糖电泳呈现3条谱带,分子量与理论值一致;扫描电镜观察表明,重组子在多杀性巴氏杆菌中成功表达,获得了细菌幽灵;菌落形成单位检验结果显示,多杀性巴氏杆菌遗传灭活率达到99%。为制备天然细菌外膜蛋白抗原疫苗提供技术依据。 The gene E of bacteriophage Phix174 and temperature-sensitivity control system were hybrid-ized with plasmid pPBAl100 by genetic engineering method to construct Recombinant pPBAl100-E. Re-combinant was transformed to Pasteurella multocida and lysis gene E expressed by temperature induction. Restriction endonuclease was applied tp detect Recombinant. Cell morphology of bacterial ghost of Pasteu-rella multocida was observed by scanning electron microscopy. The inactivation ratio was estimated by CFU analysis. Our results indicated that three bands were observed by restriction endonuclease and agarose elec- trophoresis,and the molecular weight of every band accord with theoretical values. SEM observations showed that recombanation plasmid expressed successfully in P. multocida and produced bacterial ghost. The results of CFU detecting demonstrated that inactivation ratio of P. multocida reached to 99 per cent. This study provided the technological basis for antigen of natural bacterial outer membrane protein prepa-ration.

关 键 词： 裂解基因 多杀性巴氏杆菌 细菌幽灵 扫描电镜

领　　域： [农业科学] [农业科学] [农业科学]