title: Restriction generated oligonucleotides utilizing the two base recognition endonuclease CviJI*.
abstract: The conversion of an anonymous DNA sample into numerous oligonucleotides is enzymatically feasible using an unusual restriction endonuclease, CviJI. Depending on reaction conditions, CviJI is capable of digesting DNA at a two or three base recognition sequence. CviJI normally cleaves RGCY sites between the G and C to leave blunt ends. Under 'relaxed' conditions CviJI* cleaves RGCY, and RGCR/YGCY, but not YGCR sites. In theory, CviJI* restriction of pUC19 (2686 bp) should produce 157 fragments, 75% of which are smaller than 20 bp. Instead, 96% of the CviJI* fragments were 18-56 bp long and none of the fragments were smaller than 18 bp. Thermal denaturation of these fragments generates sequence specific oligonucleotides homologous for the cognate template. The enzymatic conversion of anonymous DNA into sequence specific oligomers has implications for several conventional and novel molecular biology procedures.
authors.affiliation: CHIMERx, Madison, WI 53704.
authors.forename: N
authors.initials: N
authors.lastname: Swaminathan
authors.forename: D
authors.initials: D
authors.lastname: George
authors.forename: K
authors.initials: K
authors.lastname: McMaster
authors.forename: J
authors.initials: J
authors.lastname: Szablewski
authors.forename: J L
authors.initials: JL
authors.lastname: Van Etten
authors.forename: D A
authors.initials: DA
authors.lastname: Mead
mesh_terms: D001483:Base Sequence; D003016:Clostridium perfringens / Q000235:genetics; D004247:DNA / Q000378:metabolism*; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism*; D006801:Humans; D016982:Micrococcus luteus / Q000235:genetics; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000378:metabolism; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction
chemical_list: D009838:Oligodeoxyribonucleotides; D015345:Oligonucleotide Probes; D004247:DNA; C075913:endodeoxyribonuclease CviJI; D015252:Deoxyribonucleases, Type II Site-Specific