Elahwany M D
Section of Microbiology, Botany Department, Faculty of Science, Alexandria University, Alexandria, Egypt.
Acta Microbiol Immunol Hung. 2007 Dec;54(4):413-20. doi: 10.1556/AMicr.54.2007.4.7.
Cloning of the oriT region derived from Lactococcus lactis subsp. lactis into plasmids is expected to convert them into mobilizable plasmids in a suitable genetic background. In some cases, however, the cloning of oriT causes deleterious DNA rearrangements. In the work presented, oriT was cloned into two temperature sensitive plasmids, pJRS290 and pUCB3522. The plasmids were transformed into E. coli and electroporated into L. lactis. Restriction digestion of the plasmids derived from the cloned transformed cells showed fragment sizes different from those expected, indicating that rearrangements had occurred. Furthermore, the inability to successfully transform E. coli with one of the plasmids supports this conclusion.
将源自乳酸乳球菌乳酸亚种的oriT区域克隆到质粒中，有望在合适的遗传背景下将它们转化为可移动质粒。然而，在某些情况下，oriT的克隆会导致有害的DNA重排。在本研究中，oriT被克隆到两个温度敏感质粒pJRS290和pUCB3522中。这些质粒被转化到大肠杆菌中，并通过电穿孔导入乳酸乳球菌。对源自克隆转化细胞的质粒进行限制性酶切，结果显示片段大小与预期不同，表明发生了重排。此外，其中一个质粒无法成功转化大肠杆菌也支持了这一结论。
