Newnham P J, Taylor D E
Department of Medical Microbiology and Infectious Diseases, University of Alberta, Edmonton, Canada.
Plasmid. 1990 Mar;23(2):107-18. doi: 10.1016/0147-619x(90)90029-c.
This study was undertaken to establish a transfer complementation system for IncH plasmids and to locate regions of incompatibility within the HI1 plasmid, R27. Two regions of R27 were found to contribute to incompatibility as determined by incompatibility testing with fragments of R27 cloned in cosmid vectors. One of these regions hybridized with the IncHI1 rep probe (Couturier et al., Microbiol. Rev. 52, 375-395, 1988). Complementation analysis was carried out using transfer-deficient mutants of R27 in combination with pHH1508a. Cosmid vectors, which contained cloned restriction fragments of R27, were able to complement selected R27 Tra- mutants, enabling the transfer-deficient plasmid to transfer at near-normal frequencies. Complementation of R27 Tra- plasmids by pHH1508a at both 26 and 37 degrees C was shown to occur, but was host-dependent in its degree. These results suggest that the transfer mechanisms of IncHI and IncHII plasmids are related.
本研究旨在建立IncH质粒的转移互补系统，并确定HI1质粒R27内的不相容区域。通过用克隆于黏粒载体的R27片段进行不相容性测试，发现R27的两个区域与不相容性有关。其中一个区域与IncHI1 rep探针杂交（Couturier等人，《微生物学评论》52，375 - 395，1988）。使用R27的转移缺陷型突变体与pHH1508a进行互补分析。含有克隆的R27限制性片段的黏粒载体能够互补选定的R27 Tra-突变体，使转移缺陷型质粒能够以接近正常的频率转移。结果表明，pHH1508a在26℃和37℃下均可对R27 Tra-质粒进行互补，但互补程度依赖于宿主。这些结果表明IncHI和IncHII质粒的转移机制相关。
