Ⅰ型鸭疫里氏杆菌基因组质粒表达文库的构建

Construction of Genomic DNA Expression Library of Riemerella anatipestifer

提取Ⅰ型鸭疫里氏杆菌的基因组DNA，以睁nI进行酶切，回收、纯化0．5—6kb之间的DNA片段，与经同样酶切并去磷酸化的质粒载体pQE30连接，转化BL21（DE3）感受态细胞，从而构建了Ⅰ型鸭疫里氏杆菌基因组的质粒表达文库，经试验表明其库容量为12000个。随机挑选的阳性克隆提取质粒酶切后证明有外源DNA片段的插入，该文库为应用体内诱导抗原技术筛选鸭疫里氏杆菌的体内诱导基因奠定了基础。

The genomic DNA of Riemerella anatipestifer serotype I was digested with Kpn Ⅰ and 0.5 - 6kb fragments were recovered. The digested DNA fragments were ligated to the linear pQE30 and used to be transformed into competent Escherichia coli BL21 （ DE3 ）, respectively. The construction of genomie DNA expression library of Riemerella anatipestifer serotype Ⅰ was finished. The number of bacterial reeombinants obtained was about 12 000. It was proved by enzyme digestion test that random samples of the positive clones contained exotic DNA fragments. This study shows that the construction of genomie DNA expression library of Riemerella anatipestifer serotype I will be beneficial for screening in - vivo - induced virulent genes of Riemerella anatipestifer by in vivo induced antigen technology .