Shanghai Veterinary Research Institute, the Chinese Academy of Agricultural Sciences, 518 Ziyue Road, Shanghai 200241, China.
J Microbiol Methods. 2013 Nov;95(2):262-7. doi: 10.1016/j.mimet.2013.09.011. Epub 2013 Sep 21.
Riemerella anatipestifer causes epizootic infectious disease in poultry and serious economic losses, especially to the duck industry. Four complete genome sequences of R. anatipestifer strains are now available. However, functional studies have been limited by the lack of an effective shuttle vector. In this study, we constructed a shuttle vector, pRES, which was able to transfer plasmid DNA between Escherichia coli and R. anatipestifer strains. The vector contains the putative replication origin from R. anatipestifer plasmid pRA7026 and a ColE1 ori for replication in R. anatipestifer and E. coli respectively. In addition, it contains oriT for transferring the vector into R. anatipestifer by conjugation, and the putative promoter of the streptothricin resistance gene of plasmid pRA0726 for heterologous gene expression in R. anatipestifer. The vector pRES will be useful in the investigation of gene function in R. anatipestifer.
鸭疫里默氏杆菌引起家禽的爆发性传染病，造成严重的经济损失，尤其对养鸭业的打击最大。目前已经有四个完整的鸭疫里默氏杆菌菌株的基因组序列，但由于缺乏有效的穿梭载体，其功能研究受到限制。在本研究中，我们构建了一个穿梭载体 pRES，该载体能够在大肠杆菌和鸭疫里默氏杆菌菌株之间转移质粒 DNA。该载体包含来自鸭疫里默氏杆菌质粒 pRA7026 的假定复制原点和 ColE1 ori，分别用于在鸭疫里默氏杆菌和大肠杆菌中的复制。此外，它还包含 oriT，用于通过接合将载体转移到鸭疫里默氏杆菌中，以及质粒 pRA0726 中链霉素抗性基因的假定启动子，用于在鸭疫里默氏杆菌中进行异源基因表达。载体 pRES 将有助于研究鸭疫里默氏杆菌中的基因功能。
