de Bruijn F J, Stroke I L, Marvel D J, Ausubel F M
EMBO J. 1983;2(10):1831-8. doi: 10.1002/j.1460-2075.1983.tb01666.x.
Multicopy plasmids containing the hisDG region of Klebsiella pneumoniae were mutagenized with transposon Tn5. The resulting plasmids were examined for their ability to complement hisD and hisG mutations in Escherichia coli. The physical location of Tn5 on each of the hisD::Tn5 and hisG::Tn5 plasmids was determined by restriction endonuclease analysis. By combining the two types of data, a precise correlated physical and genetic map of the K. pneumoniae hisDG region was constructed. Based on this analysis, the minimum sizes of the hisD and the hisG genes were calculated to be 1100 bp and 900 bp, respectively. The hisO(P) region was also identified. The insertional specificity of transposon Tn5 was shown to be very low. One unanticipated result was obtained: Tn5 insertions in the plasmid-borne hisG gene were not polar on hisD.
含有肺炎克雷伯菌hisDG区域的多拷贝质粒用转座子Tn5进行诱变。检测所得质粒在大肠杆菌中互补hisD和hisG突变的能力。通过限制性内切酶分析确定Tn5在每个hisD::Tn5和hisG::Tn5质粒上的物理位置。结合这两类数据，构建了肺炎克雷伯菌hisDG区域精确的物理和遗传相关图谱。基于此分析，计算出hisD和hisG基因的最小大小分别为1100 bp和900 bp。还鉴定出了hisO(P)区域。结果表明转座子Tn5的插入特异性非常低。获得了一个意外结果：质粒携带的hisG基因中的Tn5插入对hisD没有极性效应。
