Mullings R, Bennett S P, Brown N L
Department of Biochemistry, School of Medical Sciences, University of Bristol, U.K.
Gene. 1988 Dec 25;74(1):245-51. doi: 10.1016/0378-1119(88)90297-1.
We have dissected the cloned PstI M and R genes to make DNA hybridization probes spanning most of the sequence. These subclones, and also the intact sequence, were used to search for nucleic acid homology by Southern blot in the DNA from twelve organisms which produce PstI isoschizomers. One of these probes, a 206-bp fragment from the N-terminal domain of the endonuclease, showed significant hybridisation in four strains (Escherichia coli strains RFL48, RFL49 and RFL83, and Streptomyces albus P). No significant hybridisation was detected with other parts of the PstI sequences. We have used computer similarity searches to look for homology between the PstI proteins and the known sequences of other type-II systems that recognise different sites. We postulate a possible recognition domain within the M.PstI methyltransferase based on similarity to the M.PaeR7 and M.TaqI methyltransferases.
我们已对克隆的PstI M和R基因进行了剖析,以制备覆盖大部分序列的DNA杂交探针。这些亚克隆以及完整序列,被用于通过Southern印迹法在十二种产生PstI同裂酶的生物体的DNA中寻找核酸同源性。其中一个探针,即来自核酸内切酶N端结构域的206bp片段,在四种菌株(大肠杆菌菌株RFL48、RFL49和RFL83,以及白色链霉菌P)中显示出显著的杂交信号。用PstI序列的其他部分未检测到明显的杂交信号。我们利用计算机相似性搜索来寻找PstI蛋白与识别不同位点的其他II型系统的已知序列之间的同源性。基于与M.PaeR7和M.TaqI甲基转移酶的相似性,我们推测了M.PstI甲基转移酶内一个可能的识别结构域。