Fedorova N D, Highlander S K
Department of Microbiology and Immunology, Baylor College of Medicine, Houston, TX 77030, USA.
Gene. 1997 Feb 28;186(2):207-11. doi: 10.1016/s0378-1119(96)00704-4.
New cloning and expression vectors that replicate both in Pasteurella haemolytica and in Escherichia coli were constructed based on a native sulfonamide (SuR) and streptomycin (SmR) resistant plasmid of P. haemolytica called pYFC1. Each shuttle vector includes an MCS and a selectable antibiotic resistance marker that is expressed in both organisms. Plasmid pNF2176 carries the P. haemolytica ROB-1 beta-lactamase gene (blaP, ApR) and pNF2214 carries the Tn903 aph3 kanamycin resistance (KmR) element. The expression vector, pNF2176, was created by placing the MCS downstream of the sulfonamide gene promoter (PsulII) on pYFC1; this was used to clone and express the promoterless Tn9 chloramphenicol resistance gene (cat, CmR) in P. haemolytica (pNF2200). A promoter-probe vector (pNF2283) was constructed from pNF2200 by deleting PsulII.
基于溶血巴斯德氏菌一种名为pYFC1的天然磺胺（SuR）和链霉素（SmR）抗性质粒，构建了可在溶血巴斯德氏菌和大肠杆菌中复制的新型克隆和表达载体。每个穿梭载体都包含一个多克隆位点（MCS）和一个在两种生物体中均能表达的选择性抗生素抗性标记。质粒pNF2176携带溶血巴斯德氏菌ROB - 1β - 内酰胺酶基因（blaP，ApR），pNF2214携带Tn903 aph3卡那霉素抗性（KmR）元件。表达载体pNF2176是通过将MCS置于pYFC1上磺胺基因启动子（PsulII）的下游构建而成；它被用于在溶血巴斯德氏菌中克隆和表达无启动子的Tn9氯霉素抗性基因（cat，CmR）（pNF2200）。通过缺失PsulII，从pNF2200构建了一个启动子探针载体（pNF2283）。
