Sanderson K E, Sciore P, Liu S L, Hessel A
Department of Biological Sciences, University of Calgary, Alberta, Canada.
J Bacteriol. 1993 Dec;175(23):7624-8. doi: 10.1128/jb.175.23.7624-7628.1993.
Locations of six Tn10s, closely linked to each of the six IS200s on the genomic cleavage map of Salmonella typhimurium LT2, were determined by digestion with XbaI and BlnI and separation of the fragments by pulsed-field gel electrophoresis; the locations were then further defined by P22-mediated joint transduction. The orientation of each IS200 with respect to its linked Tn10 was determined by Southern blotting. The locations of IS200-I, IS200-III, and IS200-V were confirmed to be close to sufD, melB, and purC, as previously indicated. IS200-II is jointly transduced with cysG. IS200-IV is near fliA; the linked Tn10 is inserted in fli, making the strain nonmotile. IS200-VI is jointly transduced with aspC but not with aroA. IS200 is transposed to a seventh site in some strains, while remaining in the other six locations described above. These data indicate that genome analysis by pulsed-field gel electrophoresis can locate the positions of Tn10s with accuracy sufficient to predict P22-mediated joint transduction.
通过用XbaI和BlnI酶切并利用脉冲场凝胶电泳分离片段,确定了鼠伤寒沙门氏菌LT2基因组切割图谱上与六个IS200紧密连锁的六个Tn10的位置;然后通过P22介导的联合转导进一步确定这些位置。通过Southern印迹法确定每个IS200相对于其连锁的Tn10的方向。如前所述,IS200-I、IS200-III和IS200-V的位置被证实靠近sufD、melB和purC。IS200-II与cysG共同转导。IS200-IV靠近fliA;连锁的Tn10插入到fli中,使菌株无运动性。IS200-VI与aspC共同转导,但不与aroA共同转导。在一些菌株中,IS200转座到第七个位点,而在其他六个上述位置保持不变。这些数据表明,通过脉冲场凝胶电泳进行基因组分析能够准确地定位Tn10的位置,足以预测P22介导的联合转导。