Rapid sequencing of cloned DNA using a transposon for bidirectional priming: sequence of the Escherichia coli K-12 avtA gene.
Rapid sequencing of cloned DNA using a transposon for bidirectional priming: sequence of the Escherichia coli K-12 avtA gene.
复制标题
使用转座子进行双向启动对克隆 DNA 进行快速测序：大肠杆菌 K-12 avtA 基因的序列。
DOI：
10.1093/nar/15.22.9461
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发表时间：
1987
影响因子：
14.9
通讯作者：
Berg,CM
中科院分区：
文献类型：
--
作者：
Liu,L;Whalen,W;Das,A;Berg,CM
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. TheavtA(alanine-valine transaminase) gene ofE. coliK-12 was sequenced by this approach. Surprisingly, gamma-delta insertions downstream of the coding region were found to significantly reduceavtAexpression. We suggest that these nondisruptive insertions probably change the DNA topology and thereby alter gene expression.
