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An insertion of Escherichia coli transposable element IS1K into the site immediately before tetracycline-resistance determinant of Bacillus subtilis chromosomal DNA fragment in cloning in E. coli
Authors:Hitoshi Amano  Reiko Sakaguchi  Kazuo Shishido
Institution:Department of Life Science, Tokyo Institute of Technology, Yokohama, Japan.
Abstract:In cloning in Escherichia coli C600 of a 4.5-kbp HindIII DNA fragment with the tetracycline-resistance determinant (tetBS908) from Bacillus subtilis GSY908 chromosome using a plasmid vector, a 5.2-kbp HindIII DNA fragment was also isolated at a ratio of 2 to 89. The two independently obtained 5.2-kbp fragments were an insertion derivative of the 4.5-kbp fragment and carried E. coli transposable element ISlK, which was inserted at the same site immediately before tetBS908 in the same direction. For the ISlK insertions, the 8-bp sequence CAAATTTT was used as a target, this having no similarity to any published sequences.
Keywords:IS1K insertion  IS1 Variant  Tetracycline-resistance gene  Chromosomal DNA              Escherichia coli                        Bacillus subtilis
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