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B6db references: 88067768

type Journal Article
authors Liu, L.; Whalen, W.; Das, A.; Berg, C. M.
title Rapid sequencing of cloned DNA using a transposon for bidirectional priming: sequence of the Escherichia coli K-12 avtA gene
journal Nucleic Acids Res
ui 88067768
year (1987)
volume 15
number 22
pages 9461-9.
keywords Amino Acid Sequence
abstract A new approach to determining the sequence of cloned DNA is described. Unique regions near each end of the transposable element gamma-delta provide a pair of "portable" primer-specific sites for bidirectional sequencing by the dideoxy chain termination method. A set of gamma- delta insertions positioned about 200 bp apart over the entire cloned DNA allowed us to determine the sequence of both strands in a single parental plasmid without subcloning. The avtA (alanine-valine transaminase) gene of E. coli K-12 was sequenced by this approach. Surprisingly, gamma-delta insertions downstream of the coding region were found to significantly reduce avtA expression. We suggest that these nondisruptive insertions probably change the DNA topology and thereby alter gene expression.
last changed 2002/11/02 00:35

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