Kröger M, Blum E, Deppe E, Düsterhöft A, Erdmann D, Kilz S, Meyer-Rogge S, Möstl D
Institut für Mikrobiologie and Molekularbiologie, Justus-Liebig-Universität Giessen, Germany.
Gene. 1995 May 19;157(1-2):43-7. doi: 10.1016/0378-1119(94)00779-r.
We have characterized a family of related restriction-modification (R-M) systems from the soil bacterium Herpetosiphon giganteus (Hgi). A comparison of their genetic organization reveals two types of regulatory proteins, called controlling ORF C. While one of these small reading frames derived from RM.HgiCI seems to be an enhancer of its own promoter, evidence is provided for a silencer function of the other ORF C derived from the closely related AvaII-type systems RM.HgiBI/CII/EI. The respective silencer function is detected during our various attempts to clone three isoschizomers with unusually high differences in their specific activity. Sequencing and site-directed mutagenesis revealed just two amino acids as being responsible for a massive increase in specific activity of these endonucleases.
我们已经对来自土壤细菌巨大吸管菌(Hgi)的一系列相关限制-修饰(R-M)系统进行了表征。对它们的基因组织进行比较后发现了两种类型的调节蛋白,称为控制开放阅读框C。虽然源自RM.HgiCI的这些小阅读框之一似乎是其自身启动子的增强子,但有证据表明源自密切相关的AvaII型系统RM.HgiBI/CII/EI的另一个ORF C具有沉默子功能。在我们多次尝试克隆三种特异性活性差异异常高的同裂酶的过程中检测到了各自的沉默子功能。测序和定点诱变表明,只有两个氨基酸导致这些内切核酸酶的特异性活性大幅增加。