Sepcific fragmentation of DNA heteroduplex molecules of two bacteriophage lambda mutants with endonuclease Si from Aspergillus oryzae.
Heteroduplex DNA molecules of two bacteriophage mutants (lambda b2 and lambda i434ct68) were obtained by the method of molecular hybridization. These heteroduplexes possessed two types of loops formed as a result of: a) deletion in one of the DNA strands; and b) substitution of a DNA fragment for nonhomological one. The digestion of heteroduplexes with single-stranded specific nuclease SI from Aspergillus oryzae produced two fragments at 37 degrees C and three ones at 55 degrees C. The separation of fragments and determination of their molecular weight were carried out by means of electrophoresis in agarose. The molecular weights both measured and preliminarily calculated proved to be close. One of the fragments was identificated by its biological activity in CaCl2-dependent infectious system with helperphage.
D001236:Aspergillus oryzae / Q000201:enzymology; D002845:Chromatography; D003090:Coliphages / Q000032:analysis*; D004279:DNA, Viral / Q000302:isolation & purification; D003851:Deoxyribonucleases / Q000378:metabolism; D004553:Electric Conductivity; D004591:Electrophoresis, Polyacrylamide Gel; D004720:Endonucleases / Q000378:metabolism*; D006882:Hydroxyapatites; D008970:Molecular Weight; D009691:Nucleic Acid Denaturation; D009693:Nucleic Acid Hybridization
D004279:DNA, Viral; D006882:Hydroxyapatites; D003851:Deoxyribonucleases; D004720:Endonucleases