Restriction sites that are blocked by Dam or Dcm methylation can be un-methylated by cloning your DNA into a dam–, dcm– strain of E. coli, such as dam–/dcm– Competent E. coli (NEB #C2925). Restriction sites can also be blocked if an overlapping site is present. In this case, ...
Kolek J, Sedlar K, Provaznik I, Patakova P. Dam and Dcm methylations prevent gene transfer into Clostridium pasteurianum NRRL B-598: development of methods for electrotransformation, conjugation, and sonoporation. Biotechnol Biofuels 2016;9:14....
缺失甲基转移酶的化学感受态大肠杆菌细胞,适合扩增无 Dam 和 Dcm 甲基化修饰的质粒。请注意,不建议将 dam– 菌株作为初级克隆/连接的宿主细胞。dam 突变会导致细胞内突变率增加和转化效率降低。DNA 应保持在 dam+ 菌株中,除非对无 Dam 或 Dcm 甲基化 DNA 有特定需求。 重点 扩增无 Dam 和 Dcm 甲基化修饰的...
Both Dam and Dcm methylations are detrimental for transformation of C. pasteurianum NRRL B-598. Methods for conjugation, electroporation, sonoporation, and a combined method for sono/electroporation were established for this strain. The methods described could be used for genetic improvement of this ...
ApplicationsActivity and specificity assays of restriction enzymes sensitive to Dam or Dcm methylation.Preparation of DNA molecular weight standards.Cloning.Quality ControlDNA concentration is confirmed spectrophotometrically. Identity and homogenity of DNA is confirmed by digestion by BclI and CfrI, and ...
[132].E. colipossesses four distinctmethyltransferases: Dam, Dcm, EcoK, and CcrM, of which only CcrM is essential forcell viability[133,134]. The most extensively studied, however, is theDam methylase, although this enzyme is not common in bacteria in general and is mostly limited to a ...
作者: RHJ Schlegel 摘要: A definition of the term "dam gene" is presented. It refers to the DNA adenine methylation gene of the Escherichia coli bacteria. It has the ability to methylate the Genome Analysis and Technology Core sequence, wherein Sau3A adheres to methylated a 年份: 2003 收...
Wagar EA, Safarians S, Pang M. Analysis of genomic DNA from Chlamydia trachomatis for Dam and Dcm methylation. FEMS Microbiol Lett 1992; 77:161-8.Wagar, E. A., S. Safarians, and M. Pang. 1992. Analysis of genomic DNA from Chlamydia trachomatis for Dam and Dcm methylation. FEMS ...
Expression of a 35S.GUS construct was, however, insensitive to dam methylation. dcm methylation had little if any effect on transient gene expression for any of these constructs. We provide indirect evidence that the critical sites of adenine methylation lie within sequences from the promoter ...