Chemical cross-linking of MvaI and EcoRII enzymes to DNA duplexes containing monosubstituted pyrophosphate internucleotide bond.
Gene
; 157(1-2): 187-90, 1995 May 19.
Article
en En
| MEDLINE
| ID: mdl-7607489
ABSTRACT
DNA duplexes containing a monosubstituted pyrophosphate internucleotide group, instead of a phosphodiester bond, were used as cross-linking reagent for the affinity modification of the restriction endonucleases EcoRII and MvaI (R.EcoRII and R.MvaI). An active group was introduced into the enzyme's recognition site or between the recognition site and flanking sequence. The substrate properties of such DNA duplexes were determined. Cross-linking specificity was demonstrated by competition experiments with unmodified substrate, as well as by the absence of cross-linking to an active duplex lacking a recognition site. It was shown that the nucleophilicity of the buffer solution and the presence of the enzyme cofactor Mg2+ dramatically affected the cross-linking yield.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Oligodesoxirribonucleótidos
/
ADN
/
Desoxirribonucleasas de Localización Especificada Tipo II
Idioma:
En
Revista:
Gene
Año:
1995
Tipo del documento:
Article
País de afiliación:
Rusia