The XmnI restriction-modification system: Cloning,expression, sequence organization and similarity between the R and M genes |
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| Institution: | 1. Department of Psychiatry and Behavioral Sciences, College of Medicine, Medical University of South Carolina, Charleston, SC, USA;2. Department of Neuroscience, College of Medicine, Medical University of South Carolina, Charleston, SC, USA;3. Department of Public Health Sciences, College of Medicine, Medical University of South Carolina, Charleston, SC, USA;4. Ralph H. Johnson Veterans Affairs Medical Center, Charleston, SC, USA;5. The Center on Alcoholism, Substance Abuse, and Addictions, University of New Mexico, Albuquerque, NM, USA;1. Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education and Tianjin, College of Biotechnology, Tianjin University of Science and Technology, 300457, PR China;2. Institute of Biology and Medicine, Wuhan University of Science and Technology, 430000, PR China;1. Department of Medical and Clinical Genetics, University of Helsinki, Helsinki, Finland;2. Department of Pathology, HUSLAB, University of Helsinki and Helsinki University Hospital, Helsinki, Finland;3. Genome-Scale Biology Research Program, Research Programs Unit, University of Helsinki, Finland;4. Department of Biosciences, University of Helsinki, Helsinki, Finland;5. Department of Surgery, Helsinki University Hospital, Helsinki, Finland;6. Sport and Health Sciences, University of Jyväskylä and Jyväskylä Central Hospital, Jyväskylä, Finland |
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| Abstract: | The xmnIRM genes from Xanthomonas manihotis 7AS1 have been cloned and expressed in Escherichia coli. The nucleotide (nt) sequences of both genes were determined. The XmnI methyltransferase (MTase)-encoding gene is 1861 by in length and codes for 620 amino acids (aa) (68660 Da). The restriction endonuclease (ENase)-encoding gene is 959 by long and therefore codes for a 319-aa protein (35275 Da). The two genes are aligned tail to tail and they overlap at their respective stop codons. About 4 × 104 units/g wet cell paste of R·XmnI was obtained following IPTG induction in a suitable E. coli host. The xmnIR gene is expressed from the T7 promoter. M·XmnI probably modifies the first A in the sequence, GAA(N)4TTC. The xmnIR and M genes contain regions of conserved similarity and probably evolved from a common ancestor. M·XmnI is loosely related to M·EcoRI. The XmnI R-M system and the type-I R-M systems probably derived from a common ancestor. |
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