Matsumoto-Mashimo Chiho, Guerout Anne-Marie, Mazel Didier
Unité Postulante Plasticité du Génome Bactérien, Institut Pasteur, 25, rue du Dr. Roux, 75724 Paris, France.
Res Microbiol. 2004 Jul-Aug;155(6):455-61. doi: 10.1016/j.resmic.2004.03.001.
We constructed a family of conditionally replicating plasmids, the pTX1 family, which are based on the IncPalpha oriV origin of replication that is dependent on the trfA-encoded protein. We constructed several Escherichia coli derivatives expressing trfA from different chromosomal loci, which can be transduced by phage P1 to any E. coli strain. The pTX1 plasmids also carry the oriTRP4 origin of transfer, and can be conjugated to E. coli, Vibrio cholerae and likely to a broad range of bacteria from the commonly used donor strains SM10 and S17-1, which sustain replication of the plasmids through the trfA gene carried by their integrated RP4. If TrfA is not provided in trans, these plasmids behave as suicide vectors. As such they can be used as a platform for a variety of applications such as those developed on the popular conditionally replicating plasmids carrying the oriVR6Kgamma origin of replication that is controlled by the Pi protein. Their ability to be used as efficient suicide vectors for gene disruption in V. cholerae has been demonstrated.
我们构建了一个条件复制性质粒家族，即pTX1家族，其基于依赖trfA编码蛋白的IncPα oriV复制起点。我们构建了几种从不同染色体位点表达trfA的大肠杆菌衍生物，它们可被噬菌体P1转导至任何大肠杆菌菌株。pTX1质粒还携带oriTRP4转移起点，并且可以从常用的供体菌株SM10和S17-1与大肠杆菌、霍乱弧菌以及可能的多种细菌进行接合，这些供体菌株通过其整合的RP4携带的trfA基因维持质粒的复制。如果没有反式提供TrfA，这些质粒就表现为自杀载体。因此，它们可以用作各种应用的平台，例如在携带由Pi蛋白控制的oriVR6Kγ复制起点的流行条件复制性质粒上开发的应用。它们作为霍乱弧菌基因破坏的有效自杀载体的能力已得到证实。
