High-level expression of the cloned genes encoding the subunits of and intact DNA methyltransferase, M.EcoR124.
Gene
; 112(1): 21-7, 1992 Mar 01.
Article
em En
| MEDLINE
| ID: mdl-1551595
ABSTRACT
We have cloned the genes coding for the two subunits (HsdM and HsdS) of the type-I DNA methyltransferase (MTase), M.EcoR124, into the specially constructed expression vector, pJ119. These subunits have been synthesized together as an intact MTase. We have also cloned the individual subunit-encoding genes under the control of the T7 gene 10 promoter or the lacUV5 promoter. High levels of expression have been obtained in all cases. While HsdM was found to be soluble, HsdS was insoluble. However, in the presence of the co-produced HsdM subunit, HsdS was found in the soluble fraction as part of an active MTase. We have partially purified the cloned multi-subunit enzyme and shown that it is capable of DNA methylation both in vivo and in vitro.
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Base de dados:
MEDLINE
Assunto principal:
Plasmídeos
/
Metilases de Modificação do DNA
/
Regulação Bacteriana da Expressão Gênica
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Vetores Genéticos
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Isopropiltiogalactosídeo
Idioma:
En
Ano de publicação:
1992
Tipo de documento:
Article