The recognition site of type II restriction enzyme BglI is interrupted

James A. Lautenberger, C. Thomas White, Nancy L. Haigwood, Marshall H. Edgell, Clyde A. Hutchison

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    Abstract

    The Type II restriction endonuclease BglI recognizes the interrupted DNA sequences 5′-G-C-C-N-N-N-N-N-G-G-C-. This sequence occurs at all locations in over 33 000 base pairs of DNA sequence where the enzyme was found to cut DNA and nowhere else. All six of the specified bases are essential parts of the site since all groups of five of the six bases occur in the DNA sequences tested and none of them are cut by BglI. The length of the block of intervening unspecified positions must be exactly five since all other sizes between zero and 15 occur in the DNA sequences searched and none are cut by BglI. The 5′-terminal nucleotides of BglI cleaved phage G4 replicative form DNA and plasmid pER18 DNA were compared with the DNA sequences near the BglI sites on these DNAs. These results indicated that BglI cuts within the intervening unspecified region and produces single-stranded 3′ termini that are three bases long. The BglI recognition site and cleavage points can thus be represented as follows: 5′-G-C-C-N-N-N-N-N-G-G-C-3′3′-C-G-G-N-N-N-N-N-C-C-G-5′ This study of the BglI recognition site was facilitated by the use of inexpensive microcomputers. A system of programs was developed that allowed analysis of over 33 kb of DNA sequences stored on flexible magnetic disks or audio cassettes. While these programs were generally written in the higher level language BASIC, some assembly language subroutines were utilized to reduce execution time.

    Original languageEnglish (US)
    Pages (from-to)213-231
    Number of pages19
    JournalGene
    Volume9
    Issue number3-4
    DOIs
    StatePublished - May 1980

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    Keywords

    • DNA sequence data base
    • Variable sticky ends
    • microcomputers
    • symmetrical interrupted hexanucleotide

    ASJC Scopus subject areas

    • Genetics

    Cite this

    Lautenberger, J. A., Thomas White, C., Haigwood, N. L., Edgell, M. H., & Hutchison, C. A. (1980). The recognition site of type II restriction enzyme BglI is interrupted. Gene, 9(3-4), 213-231. https://doi.org/10.1016/0378-1119(90)90324-K