|
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 |
Activity |
2.6.1.66 |
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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