炭疽芽胞杆菌nprR突变株的生物学特性分析  

Biological Characteristics of Bacillus anthracis nprR Deletion Mutant

作　　者：袁利思 陶好霞 江巍 关清 王博文 刘纯杰 刘东[1] 王艳春 YUAN Li-Si;TAO Hao-Xia;JIANG Wei;GUAN Qing;WANG Bo-Wen;LIU Chun-Jie;LIU Dong;WANG Yan-Chun(College of Life Sciences,Hebei Normal University,Shijiazhuang 050024;Beijing Institute Beijing 100071,China)

机构地区：[1]河北师范大学生命科学学院,河北石家庄050024 [2]军事科学院军事医学研究院生物工程研究所,北京100071

出　　处：《生物技术通讯》2018年第4期491-495,共5页Letters in Biotechnology

摘　　要：目的:分析炭疽芽胞杆菌npr R突变株的生物学特性,研究基因缺失对菌株的影响。方法:利用PCR、免疫印迹等方法在基因水平和蛋白水平对菌株进行鉴定分析;绘制生长曲线比较突变株和A16R株的生长性能;芽胞形成实验分析突变株形成芽胞能力,糖酵解实验分析二者生化特性的差异。结果:突变株npr R基因缺失无回复突变,与出发株A16R相比,突变株蛋白酶活性大大降低,保护性抗原降解现象明显减少,生长速度和生化特性基本一致,但芽胞形成能力明显下降。结论:与A16R株相比,突变株蛋白酶活性降低且芽胞形成能力下降,可作为一种新的抗原表达宿主菌用于相关疫苗的抗原表达与制备。Objective： To analyze nprR gene deletion mutant of Bacillus anthracis vaccine strain A16R. Methods： The recombinant strain was identified by PCR and extracellular protease activity test. Expression of protective antigen and Npr599 were determined using Western blotting. The biochemical characteristics were detected by using API 50 CH. The growth rate was analyzed by drawing one-step growth curve and the sporulation of mutant also was investigated by spore stain. Results： Compared with the A16R strain, the extracellular proteolytic enzyme activity of nmtant decreased significantly, and the expression of neutral metalloprotease Npr599 also approached to zero. The mutant strain produced protective antigen with minimal to no degradation during the growth. These two strains have the same growth rate and the biochemical characteristic, while the mutant strainfailed to initiate sporulation when grown in LB medium. Conclusion： The nprR gene deletion mutant strain is low extracellular proteolytic enzyme activity and can potentially serve as a host strain for anthrax vaccine antigen preparation.

关 键 词：炭疽芽胞杆菌 nprR基因 生物特性 蛋白酶活性 

分 类 号：Q78[生物学—分子生物学]