Open AccessPhsical Map of Phage 2C DNA: Evidence for the Existence of Large Redundant Ends
Author(s) -
CONENE Marc,
HOET Philippe,
COCITO Carlo
Publication year - 1983
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1983.tb07326.x
Subject(s) - restriction enzyme , dna , ecori , restriction fragment , restriction map , biology , genome , genetics , microbiology and biotechnology , hindiii , restriction site , base pair , gene , nucleic acid sequence
The chromosome o0f the Bacillus Subtilkis phage 2C, a linear nmolecule of duble‐stranded DNA of about 10 8 Da, in which thymine is completely replaced by hydroxymethluracil, was cleaved by different endonucleases. In some cases restriction segmentsd were much fewer than expectded, suggesting a possible interference oif the unusual base with the recognition mechanism of endonucleases. The physical map of 2C DNA was established by use fo Sal I and Hae III restriction endonucleases, which yielded a limited numbver of fragments. The expected number of fragments was 240 for Hae III and 23 for Sal I; in realitym, five segmenbts were obsereved up[on clevage with Hae III and four with Sal I. The terminal fragments of the genome were first identified; the other fragments were ordered by hybridizaiton and maoleculkar wight determination of restriction fragments showed the prewsence of homologous regfions at the ends of the 2C genome. The structure fo thses direct repretitive sequences was analyzed by clevage with Hae III and hybridization with Eco RI restriction fragments. Their size (9.2 MDa) was found to be about 1/11 of that of the whole chromnosome.