Cooperative Binding Properties of Restriction Endonuclease EcoRII with DNA Recognition Sites
نویسندگان
چکیده
منابع مشابه
Asymmetric photocross-linking pattern of restriction endonuclease EcoRII to the DNA recognition sequence.
The EcoRII homodimer engages two of its recognition sequences (5'-CCWGG) simultaneously and is therefore a type IIE restriction endonuclease. To identify the amino acids of EcoRII that interact specifically with the recognition sequence, we photocross-linked EcoRII with oligonucleotide substrates that contained only one recognition sequence for EcoRII. In this recognition sequence, we substitut...
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the enzyme DpnII, produced by another strain of D. pneumoniae, recognizes the same sequence, but is inhibited by this same methylated base. The significance of the occurrence of modified nucleotides in eukaryotic DNA is not as yet understood, although a relationship between modification and gene expression has been proposed by several workers (Waalwijk & Flavell, 1978; Bird et al., 1979). By us...
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The role of particular residues of the PvuII endonuclease in DNA binding and cleavage was studied by mutational analysis using a number of in vivo and in vitro approaches. While confirming the importance of residues predicted to be involved directly in function by the crystal structure, the analysis led to several striking results. Aspartate 34, which contacts the central base pair of the PvuII...
متن کاملDigestion of terminal restriction endonuclease recognition sites on PCR products.
One of the common methods for cloning polymerase chain reaction (PCR) products is overhanging-end cloning (also known as sticky-end or directional cloning). Frequently, it is not possible to use restriction enzyme sites already present in the amplified product, and primers that encode recognition sites of restriction endonucleases in addition to the specific sequence have to be designed. After ...
متن کاملEfficient cloning of PCR generated DNA containing terminal restriction endonuclease recognition sites.
DNA generated by the polymerase chain reaction (PCR) (1) containing terminal restriction endonuclease recognition sites to permit cloning usually relies on the use of unphosphorylated primers incorporating a restriction endonuclease recognition site of choice plus 3 —4 extra 5' bases flanking that site. Various sites (Notl, Xhol and Xbal (2), for example) incorporated into the termini of PCR pr...
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ژورنال
عنوان ژورنال: Journal of Biological Chemistry
سال: 1998
ISSN: 0021-9258
DOI: 10.1074/jbc.273.14.8294