Iodouracil-mediated photocrosslinking of DNA to EcoRII restriction endonuclease in catalytic conditions
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...
متن کاملCatalytic and DNA binding properties of PvuII restriction endonuclease mutants.
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 کاملRestriction endonuclease analysis of Aspergillus fumigatus DNA.
AIMS To develop a genome based DNA fingerprinting system for Aspergillus fumigatus mould. METHODS DNA was extracted from 21 isolates obtained from eight patients with an aspergilloma. This was with a freeze-dried mycelial extract fragmented in liquid nitrogen. DNA was subsequently purified by phenol-chloroform extraction followed by ultracentrifugation on a caesium chloride gradient. The DNA ...
متن کاملEndonuclease-mediated long PCR and its application to restriction mapping.
The polymerase chain reaction (PCR) is the most widely used technique for the study of DNA. Applications for PCR have been extended significantly by the development of "long" PCR, a technique that makes it possible to amplify DNA fragments up to 40 kb in length. This article describes two novel applications of the long PCR technique, one which simplifies restriction mapping and another which en...
متن کاملUnexpected structural diversity in DNA recombination: the restriction endonuclease connection.
Transposition requires a coordinated series of DNA breakage and joining reactions. The Tn7 transposase contains two proteins: TnsA, which carries out DNA breakage at the 5' ends of the transposon, and TnsB, which carries out breakage and joining at the 3' ends of the transposon. TnsB is a member of the retroviral integrase superfamily whose hallmark is a conserved DDE motif. We report here the ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: Photochemical & Photobiological Sciences
سال: 2002
ISSN: 1474-905X,1474-9092
DOI: 10.1039/b202028a