Genomic replacement in Escherichia coli K-12 using covalently closed circular plasmid DNA.

Genomic replacement in Escherichia coli K-12 using covalently closed circular plasmid DNA.
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使用共价闭合环状质粒 DNA 在大肠杆菌 K-12 中进行基因组替换。

DOI:
10.1016/0378-1119(90)90337-q
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发表时间:
1990
期刊:
影响因子:
3.5
通讯作者:
Gennis,RB
Gennis,RB
中科院分区:
生物学3区
文献类型:
--
作者:
Oden,KL;DeVeaux,LC;Vibat,CR;CronanJr,JE;Gennis,RB

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用共价闭合环状质粒DNA在大肠杆菌(JC 7623)的herecB_(21)recC_(22)sbcB_(15)asbcC_(201)突变体中构建了多个不同位点的基因置换。通过Southern印迹证实了代表由涉及ccc质粒分子和基因组的双交换事件形成的缺失和插入突变的所选构建体,并评估了双交换事件的频率。据报道,如前所述,这种突变体可以在不线性化质粒DNA的情况下构建。
A number of gene replacements at different loci were constructed using covalently closed circular (ccc) plasmid DNA in herecB21recC22sbcB15asbcC201mutant ofEscherichia coli(JC7623). Selected constructs representing deletions and insertion mutations formed from double-crossover events involving the ccc plamid molecules and the genome were confirmed by Southern blots, and the frequency of double-crossover events was evaluated. It is reported that such mutants may be constructed without linearizing plasmid DNA, as described previously.
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