牦牛副伤寒沙门氏菌特异性抗原基因在大肠杆菌中的克隆及表达  

CLONING AND EXPRESSION OF THE SPECIFIC ANTIGENIC GENE OF SALMONELLA MORBIFICANS(YAK PARATYPHUS) IN E. coil

作　　者：裴青生[1] 解洪业[1] 王爱萍[1] 任晓凤[1] 

机构地区：[1]青海省畜牧兽医科学院,西宁810003

出　　处：《青海畜牧兽医杂志》1992年第6期5-8,共4页Chinese Qinghai Journal of Animal and Veterinary Sciences

摘　　要：对牦牛副伤寒牛病沙门氏菌(S. morbificans bovis)的染色体DNA进行提取,用基因工程中的“鸟枪法”,将限制性内切酶部分酶切染色体DNA片段,重组到载体pBR_(322)的Barn H I酶切位点上,再转化到大肠杆菌DH_(5α)中,经耐药表型筛选,在约1900个转化菌中找出了86个具有重组质粒的细菌株,经ELISA法检测,其中pS9,pS31表现出较强的表达,通过动物免疫试验,小鼠获5/8保护,豚鼠获6/8保护,大肠杆菌DH_(5α)无保护力(0/4,0/8),证明本试验所克隆的片段是保护性抗原,为今后构建基因工程苗提供了依据。After extracting the chromosome DNA of S. morbificans, some fragments of chronlosome DNA obtained by festrictive endonuclease have been recombined to Bam H I site of carrier pBR_(322), by means of 'fowling piece test', then pBR_(322) has been transferred into E. coli DH_(5α). Screening test of drug resistance showed that 86 of 1900 treated cells possessed the recombination plasmid. The ELISA test showed that pS9 and pS31 possessed the fairly good expression. Immune tests on experimental animals showed that 5/8 of mice and 6/8 of quinea pigs aquired protection, but E. coli. DH_(5α)(0/8 and 0/4)had not any protection. These results indicated that the cloning fragment in this study belong to genome of protective antigen, and it has supplied a good base for genetic engineering vaccine in future.

关 键 词：牦牛副伤寒 基因重组 基因表达 

分 类 号：S8[农业科学—畜牧兽医]