摘要
从临床病鸭中分离并鉴定了1型鸭疫里默氏杆菌，并在提取鸭疫里默氏杆菌的基因组DNA后，用Sau3A I酶切，回收大小为0．07～4 kb的片段；将酶切片段与酶切的质粒载体pRSET连接后，电转化大肠杆菌Rosetta，成功构建了1型鸭疫里默氏杆菌的基因组文库，经检测库容量约为40000个。随机筛选30个单菌落，用pRSET通用引物进行PCR鉴定，统计结果显示插入片段的大小93．3％在1～3 kb范围内，成功构建了鸭疫里默氏杆菌基因组文库，为鸭疫里默氏杆菌功能基因的克隆及鉴定奠定了基础。
Riemerella anatipestifer infection was probably the most economically important infectious disease of farm ducks worldwide. In this study, one strain of R. anatipestifer serotype 1 was isolated and identified. Then the genomic DNA of R. anatipestifer serotype 1 was extracted and digested with Sau 3A I and 0.07~4 kb fragments were recovered. The digested bacterial DNA fragments was ligated to the digested pRSET and transformed into competent Escherichia coli Rosetta. Finally, high quality genomic DNA library of R. anatipestifer serotype 1 was succeeded to be constructed, and its content was about 40000, and 93. 3% DNA inserts were from 1 to 3 kb. Further research about functional proteins could be done.
出处
《中国兽药杂志》
2014年第1期11-14,共4页
Chinese Journal of Veterinary Drug
关键词
鸭疫里默氏杆菌
分离鉴定
基因组文库
Riemerella anatipestifer
isolation and identification
genomic DNA library
