ATYPICAL DNA-BINDING PROPERTIES OF CLASS-IIS RESTRICTION ENDONUCLEASES - EVIDENCE FOR RECOGNITION OF THE COGNATE SEQUENCE BY A FOKI MONOMER

被引：35

作者：

SKOWRON, P ^{[1
]}

KACZOROWSKI, T ^{[1
]}

TUCHOLSKI, J ^{[1
]}

PODHAJSKA, AJ ^{[1
]}

机构：

[1] UNIV GDANSK,DEPT MICROBIOL,UL KLADKI 24,PL-80822 GDANSK,POLAND

来源：

GENE | 1993年 / 125卷 / 01期

关键词：

PROTEIN DNA INTERACTION; GEL-MOBILITY-SHIFT ASSAY; ENZYME; RESTRICTION-MODIFICATION SYSTEM;

D O I：

10.1016/0378-1119(93)90738-O

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

The DNA-binding properties of the FokI restriction endonuclease were studied using the gel-mobility-shift assay. Specific recognition of the cognate sequence and cleavage of DNA are distinguishable functions and can be separated. FokI binds to its recognition site predominantly as a monomer. At high concentrations, FokI exhibits a cooperative recognition sequence-dependent aggregation. In 20 mM KCl/10 mM Tris.HCl buffer, the binding constant of FokI to its cognate site is equal 6.0-7.9 x 10(8)/mol and is lower than the values for most gene-regulatory proteins. FokI binding is 600-1500 times weaker to non-cognate double-stranded DNA than to the GGATG site, and 30 000 times weaker to single-stranded DNA or tRNA. The method of Bading [Nucleic Acids Res. 16 (1988) 5241-5248], used for determining the stoichiometry of protein bound to DNA by gel-mobility-shift assay, is extended.

引用

页码：1 / 10

页数：10

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