摘要
通过将2种不同的重组溶菌质粒转化至鸡痢疾志贺菌中,构建含有双重溶菌质粒的志贺菌重组菌株,以提高该菌菌壳制备的裂解率和稳定性,并对其特性进行研究。用重组溶菌质粒pBV220::E和pBBRIMCS-E共同转化到鸡痢疾志贺菌,构建志贺菌重组菌株Sd(pBV220::E/pBBRI MCS-E),重组菌株于28℃培养至D600值为0.5时升至42℃继续培养,间隔30min检测菌液D600值,进行重组菌株的溶菌裂解动力学比较试验,绘制裂解曲线图并计算出裂解率,制备菌壳并对其形态进行电镜观察。重组菌株Sd(pBV220::E/pBBRIMCS-E)经42℃诱导后可形成菌壳且保留了基本的细胞形态,裂解率达到了99.999 9%。本研究成功制备出鸡痢疾志贺菌菌壳,为研究利用该菌菌壳预防鸡痢疾志贺菌病的可行性奠定了基础。
In order to improve lysis rate and stability of chicken origin Shigella dysenteriae ghosts,recombinant strain of Shigella with double bacteriolytic plasmids was constructed and its characteristics was studied. Two recombinant bacteriolysis plasmid pBV220：：E and pBBR IMCS- E were co-transformed into the Shigella strain isolated from chicken to produce recombinant strain Sd（pBV220：：E /pBBRIMCS-E）. The recombinant strain Sd（pBV220：： E /pBBRIMCS-E） was grown at 28℃ with shaking until mid log-phase,followed by incubation at 42℃ to induce the expression of gene E and produce Shigella dysenteriae ghosts. The D600 value of bacterial cultures was monitored over 20 h at 30 min intervals. The morphology of chicken origin Shigella dysente- riae ghosts was observed by electron microscope. The recombinant strains Sd（pBV220：E/pB- BRIMCS-E） generated chicken origin Shigella dysenteriae ghosts after inducing at 42℃. The ghosts retained basic cell morphology,with the lysis rate of 99. 999 9%. The chicken origin Shi- gella dysenteriae ghosts were generated successfully with double bacteriolysis plasmids, which laid foundation for future development of bacterial ghost vaccine against Shigella infection in chicken.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2013年第3期371-375,共5页
Chinese Journal of Veterinary Science
基金
国家自然科学基金资助项目(31272568)
关键词
志贺菌
裂解基因E
菌壳
疫苗
Shigella
lysis gene E
bacterial ghosts
vaccine
