Cloning and characterization of the genes encoding the MspI restriction modification system

Lin, P. M., Lee, C. H., Roberts, R. J. (April 1989) Cloning and characterization of the genes encoding the MspI restriction modification system. Nucleic Acids Res, 17 (8). pp. 3001-11. ISSN 0305-1048

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URL: http://www.ncbi.nlm.nih.gov/pubmed/2471145
DOI: 10.1093/nar/17.8.3001

Abstract

The genes encoding the MspI restriction modification system, which recognizes the sequence 5' CCGG, have been cloned into pUC9. Selection was based on expression of the cloned methylase gene which renders plasmid DNA insensitive to MspI cleavage in vitro. Initially, an insert of 15 kb was obtained which, upon subcloning, yielded a 3 kb EcoRI to HindIII insert, carrying the genes for both the methylase and the restriction enzyme. This insert has been sequenced. Based upon the sequence, together with appropriate subclones, it is shown that the two genes are transcribed divergently with the methylase gene encoding a polypeptide of 418 amino acids, while the restriction enzyme is composed of 262 amino acids. Comparison of the sequence of the MspI methylase with other cytosine methylases shows a striking degree of similarity. Especially noteworthy is the high degree of similarity with the HhaI and EcoRII methylases.

Item Type: Paper
Uncontrolled Keywords: Amino Acid Sequence Bacterial Proteins/genetics Base Sequence Cloning, Molecular Comparative Study DNA Restriction-Modification Enzymes/*genetics DNA, Bacterial/genetics Deoxyribonuclease HpaII Deoxyribonucleases, Type II Site-Specific/*genetics *Genes, Bacterial Molecular Sequence Data Moraxella/enzymology/*genetics Research Support, U.S. Gov't, Non-P.H.S. Restriction Mapping Site-Specific DNA Methyltransferase (Cytosine-Specific)/*genetics
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > restriction enzyme
CSHL Authors:
Communities: CSHL labs > Roberts lab
Depositing User: Gail Sherman
Date: 25 April 1989
Date Deposited: 06 Jul 2017 18:35
Last Modified: 08 Nov 2017 17:01
PMCID: PMC317708
Related URLs:
URI: http://repository.cshl.edu/id/eprint/34871

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