Boissinot M, Mercier J, Levesque R C
Antimicrob Agents Chemother. 1987 May;31(5):728-34. doi: 10.1128/AAC.31.5.728.
Cloning of a 6.3-kilobase BglII DNA fragment from plasmid R46 permitted the isolation of the OXA-2 beta-lactamase gene. Selected DNA fragments internal and adjacent to the OXA-2 beta-lactamase structural gene were used as probes in homology studies with other plasmid-mediated beta-lactamases. Under conditions of high stringency, no cross hybridization could be detected with DNA probes from within the open reading frame of the OXA-2 structural gene. At a lower stringency, one of two DNA fragments used as probes cross hybridized weakly with the OXA-3 bla gene. Other DNA fragments tested and known to contain sequences flanking the OXA-2 determinant cross hybridized with OXA-3 and PSE-4 plasmid DNA. From the known nucleotide sequence of OXA-2 and TEM-1, we synthesized a series of oligonucleotides corresponding to sequences internal to their respective structural genes. A 12-mer oligonucleotide containing the OXA-2-active-site nucleotide sequences cross hybridized only with OXA-3. All other oligonucleotides tested were found to be specific for their respective OXA-2 or TEM-1 gene. Such beta-lactamase gene probes should facilitate studies of beta-lactamase molecular epidemiology and beta-lactamase gene polymorphism.
从质粒R46中克隆出一个6.3千碱基的BglII DNA片段，使得OXA - 2β-内酰胺酶基因得以分离。选择OXA - 2β-内酰胺酶结构基因内部及相邻的DNA片段作为探针，用于与其他质粒介导的β-内酰胺酶进行同源性研究。在高严格条件下，未检测到与OXA - 2结构基因开放阅读框内的DNA探针发生交叉杂交。在较低严格条件下，用作探针的两个DNA片段之一与OXA - 3 bla基因发生了弱交叉杂交。测试的其他已知含有OXA - 2决定簇侧翼序列的DNA片段与OXA - 3和PSE - 4质粒DNA发生了交叉杂交。根据OXA - 2和TEM - 1的已知核苷酸序列，我们合成了一系列对应于它们各自结构基因内部序列的寡核苷酸。一个含有OXA - 2活性位点核苷酸序列的12聚体寡核苷酸仅与OXA - 3发生交叉杂交。测试的所有其他寡核苷酸对它们各自的OXA - 2或TEM - 1基因具有特异性。此类β-内酰胺酶基因探针应有助于β-内酰胺酶分子流行病学和β-内酰胺酶基因多态性的研究。
