Transposon-facilitated DNA sequencing.
Transposon-facilitated DNA sequencing.
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转座子促进的 DNA 测序。
DOI：
10.1073/pnas.88.4.1247
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发表时间：
1991
影响因子：
11.1
通讯作者：
Palazzolo,MJ
中科院分区：
文献类型：
--
作者：
Strathmann,M;Hamilton,BA;Mayeda,CA;Simon,MI;Meyerowitz,EM;Palazzolo,MJ
我们在此描述了一种基于转座子的 DNA 测序策略，该策略允许通过细菌交配在整个目标序列中引入测序引发位点。设计了小质粒来选择转座子插入载体。通过聚合酶链反应和提供模板的细菌过夜培养物来绘制转座子插入位点。然后可以对一小组具有数百个碱基对间隔的转座子的质粒进行测序。对应于转座子末端的测序引物允许双向测序。因此，可以很容易地确定两条链的整个序列。
We describe here a transposon-based DNA sequencing strategy that allows the introduction of sequencing priming sites throughout a target sequence by bacterial mating. A miniplasmid was designed to select against transposon insertions into the vector. Sites of transposon insertion are mapped by the polymerase chain reaction with bacterial overnight cultures providing the templates. A small set of plasmids with transposons spaced several hundred base pairs apart can then be sequenced. Sequencing primers corresponding to the transposon ends allow sequencing in both directions. Thus, the entire sequence of both strands can be easily determined.