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Nucleic Acids Research, 1981, Vol. 9, No. 21 5679-5688
© 1981

MOLECULAR BIOLOGY

Specific deletion of DNA sequences between preselected bases

Nikos Panayotatos and Kim Truong

Biogen S.A., 3, route de Troinex, 1227 Carouge/Geneva, Switzerland

Received September 18, 1981. Blunt-end ligation of a "filled-in" HindIII, Sal I, Ava I or Bcl I restriction site with a DNA fragment having A, G, C, or Tas the terminal 3' nucleotide regenerates the corresponding restriction site. A combination of this property with the action of BAL 31 nuclease which progressively removes base-pairs from the ends of linear DNA, can generate deletions extending to desired pre-selected nucleotides, and introduces unique restriction sites at those positions. Similarly other restriction sites can be used to select for the deletion of sequences between specific di-, tri-, tetra- and penta-nucleotides. Using this method, 10 base pairs were deleted from the end of a restriction fragment carrying the late promoter for bacterio phage T7 gene1.1, to create a molecule with a unique restriction site at the initiation cod on for translation.


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