Sequence, Internal Homology and High-Level Expression of the Gene for a DNA-(Cytosine N4)-Methyltransferase, M-Pvu-Ii

Tao, T., Walter, J., Brennan, K. J., Cotterman, M. M., Blumenthal, R. M. (June 1989) Sequence, Internal Homology and High-Level Expression of the Gene for a DNA-(Cytosine N4)-Methyltransferase, M-Pvu-Ii. Nucleic Acids Research, 17 (11). pp. 4161-4175. ISSN 0305-1048

URL: http://www.ncbi.nlm.nih.gov/pubmed/2662138
DOI: 10.1093/nar/17.11.4161

Abstract

The base sequence of the pvulIM gene has been determined. This gene codes for a DNA-(cytosine N4-methyltransferase, M.Pvu II. The base sequence contains a single large open reading frame that predicts a 38.3kDa polypeptide, consistent with experimental data. The pvulIM gene contains some sequences common to DNA methyltransferases in general, but includes none of the sequences specifically conserved among DNA-(cytosine 5)-methyltransferases. The pvulIM sequence also reveals an internal homology at the amino acid level, each half of which spans over 100 amino acids and is itself homologous to the sequences of some DNA-(adenine N5)-methyltransferases. A derivative of the pvulIM plasmid was constructed to allow high-level production of M.Pvu II. Specifically, the composite plac promoter was inserted 5′ to pvulIM, intervening DNA was deleted, and the resulting construct was used to transform an mcrB laclq strain of Escherichia coli. When this transformant was induced with isopropyl-B-D-galactopyranoside (IPTG), growth rapidly ceased and M.Pvu II accumulated to the point of comprising over 10% of the total soluble protein.

Item Type: Paper
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > genes, structure and function
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > methyltransferase
Communities: CSHL labs
Depositing User: Gail Sherman
Date: 12 June 1989
Date Deposited: 06 Jun 2017 15:34
Last Modified: 06 Jun 2017 15:34
PMCID: PMC317926
Related URLs:
URI: http://repository.cshl.edu/id/eprint/34912

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