Wood A R, Lainson F A, Wright F, Baird G D, Donachie W
Moredun Research Institute, Edinburgh.
Res Vet Sci. 1995 Mar;58(2):163-8. doi: 10.1016/0034-5288(95)90071-3.
The complete nucleotide sequence of a 4.3 kilobase pair plasmid, pAB2, isolated from a bovine strain of Pasteurella haemolytica serotype A1, was determined. It encodes a Rob-1 type beta-lactamase and a region with homology to the mobilisation (mob) region of the Escherichia coli plasmid, ColE1. An insertion mutant of pAB2 (pTC2/81) carrying a copy of Tn5 was transferred to E coli K12 by conjugation. Subsequently pTC2/81 could be transferred by transformation to E coli HB101, but not to P haemolytica serotypes A1 or A2. However, a derivative of this construct containing only a fragment of the Tn5 insertion sequence was able to transform P haemolytica. A further construct containing a fragment of the P haemolytica A1 leucotoxin A gene, was similarly restricted to transforming E coli. These results demonstrate that the pAB2 plasmid is capable of acting as an E coli/P haemolytica shuttle vector. However, the nature of the cloned DNA sequences are important to transformation.
从溶血巴斯德氏菌A1血清型牛源菌株中分离出一个4.3千碱基对的质粒pAB2,并测定了其完整的核苷酸序列。它编码一种Rob-1型β-内酰胺酶以及一个与大肠杆菌质粒ColE1的转移(mob)区域具有同源性的区域。携带Tn5拷贝的pAB2插入突变体(pTC2/81)通过接合转移至大肠杆菌K12。随后,pTC2/81可通过转化转移至大肠杆菌HB101,但不能转移至溶血巴斯德氏菌A1或A2血清型。然而,该构建体的一个仅包含Tn5插入序列片段的衍生物能够转化溶血巴斯德氏菌。另一个包含溶血巴斯德氏菌A1白细胞毒素A基因片段的构建体同样仅限于转化大肠杆菌。这些结果表明,pAB2质粒能够作为大肠杆菌/溶血巴斯德氏菌穿梭载体。然而,克隆的DNA序列的性质对转化很重要。