摘要
在从患病大口鲇中分离到普通变形杆菌TWN-3株的基础上,将该菌总DNA用EcoRI不完全酶切后,分离5kb左右的片段,连接入pUC18质粒,转化E.coli BL21,构建基因组文库,得到3.6×103个重组子,远大于理论计算的1831个重组子,随机挑选重组子经EcoRI酶切鉴定重组率为100%,文库构建成功;将该菌裂解液与佐剂混合,免疫雄性新西兰大白兔,免疫程序结束后,抽血并分离血清,对用异丙基-β-D-硫代半乳糖苷(IPTG)诱导后的文库进行免疫印迹筛选,最终得到8个阳性克隆子。本实验为该菌的DNA疫苗以及重要基因表达研究打下基础。
The strain of Proteus vulgaris was isolated from the catfish with disease. By identification, the strain was named TWN-3. Genomic DNA of TWN-3 was extracted and partially digested by restriction enzyme EcoRI, then ligated with plasmid pUC18 . After transforming E. coli BL21, the genomic library was constructed and 3.6×10^3 recombinants were obtained. The results after digested by restriction enzyme EcoRI revealed that the recombinants which had been randomly picked out were all contained foreign DNA fragments. The ratio of the recombinants was 100%. The results suggested that the gene library was constructed successfully. Eight positive recombinants were screened from the gene library by using rabbit anti- Proteus vulgaris TWN-3 strain differential proteins serum.
出处
《淡水渔业》
CSCD
北大核心
2007年第2期12-15,共4页
Freshwater Fisheries
基金
通威科技发展项目
四川省科技攻关项目(05NG002-002-4)
