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Cloning of the resistant Eco RII recognition site of phage T7 into an Eco RII‐sensitive plasmid makes the site susceptible to the restriction enzyme
Author(s) -
Krüger Detlev H.,
Prösch Susanna,
Reuter Monika,
Goebel Werner
Publication year - 1990
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.3620300913
Subject(s) - restriction enzyme , plasmid , pbr322 , recognition sequence , bacteriophage , recombinant dna , restriction site , cloning (programming) , dna , biology , endonuclease , genetics , cloning vector , microbiology and biotechnology , gene , escherichia coli , vector (molecular biology) , computer science , programming language
The recognition sequence 5'‐CC(A/T)GG for Eco RII in the bacteriophage T7 genome is refractory to this restriction endonuclease, despite not bearing the specific (protective) methylation. Following the integration of this site as part of a 219 bp fragment (in which the recognition sequence is flanked by about 100 bp of T7 origin) into the Eco RII‐sensitive vector pUC18, the T7 site becomes susceptible to cleavage, too. The same is true of recombinant pBR322 plasmids containing the T7‐derived recognition site. The results show that the flanking sequences are not immediately responsible for the refractory behaviour of Eco RII sites and are in agreement with data according to which Eco RII requires the coordinated presence of at least two recognition sites in its DNA substrate.