[DNA-like duplexes containing repetitive sequences. VIII. Synthesis and properties of DNA fragments--substrates of restriction endonuclease EcoRII].
The optimal way of constructing a family of restriction endonuclease EcoRII substrates has been developed. The substrates are DNA-like duplexes containing regularly repeated native or modified sites of this enzyme as well as those of EcoRI and AluI. Synthesis of substrates was performed by water-soluble carbodiimide-induced polycondensation of two nonanucleotides, d(C-C-T-G-G-A-A-T-Tp) and d(C-C-A-G-G-A-G-C-Tp), as constituents of different complementary complexes. The products of reaction (degree of polymerization, 2-20) were isolated by G-200 gel-filtration. The yield of polymers was about 70%. The main products of reaction were dimers when dephosphorylated nonanucleotides (terminators of polycondensation) were used. The thermal stability of DNA-like duplexes is very high. The structure of the polymers obtained has been confirmed by UV-spectroscopy and by CD data as well as by the results of cleavage by EcoRI and AluI restriction endonucleases.
D001483:Base Sequence; D002850:Chromatography, Gel; D002942:Circular Dichroism; D004262:DNA Restriction Enzymes / Q000235:genetics*; D004269:DNA, Bacterial / Q000032:analysis / Q000235:genetics*; D015252:Deoxyribonucleases, Type II Site-Specific; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D009692:Nucleic Acid Heteroduplexes / Q000032:analysis; D009841:Oligonucleotides / Q000032:analysis / Q000138:chemical synthesis*; D012091:Repetitive Sequences, Nucleic Acid; D013379:Substrate Specificity
D004269:DNA, Bacterial; D009692:Nucleic Acid Heteroduplexes; D009841:Oligonucleotides; D004262:DNA Restriction Enzymes; C046545:CCWGG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific
DNK-podobnye dupleksy, soderzhashchie povtory. VIII. Sintez i svoĭstva fragmentov DNK--substratov éndonukleazy restriktsii EcoRII.