An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis.
An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis.
复制标题
通过寡核苷酸介导的诱变构建的改进的正选择质粒载体。
DOI：
10.1093/nar/11.22.8019
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发表时间：
1983
影响因子：
14.9
通讯作者：
L. Philipson
中科院分区：
文献类型：
--
作者：
B. Nilsson;M. Uhlén;S. Josephson;S. Gatenbeck;L. Philipson
构建了一种大肠杆菌质粒载体pUN121，它允许对携带DNA插入物的转化子进行阳性选择。携带DNA插入子的转化子的阳性选择。载体基于质粒pTR262 （Roberts et al.）。基因，12,(1980),123-127)，其中四环素耐药基因受噬菌体lambda cI基因编码的抑制蛋白的转录控制。利用体外重组技术（包括寡核苷酸介导的诱变）对该质粒进行了重排，得到了一个更小的质粒（4.4 kb），除了具有独特的HindIII和BclI位点外，还具有EcoRI、XmaI和SmaI的独特克隆位点。质粒具有一个功能性氨苄西林耐药基因，利用新的限制性位点（EcoRI、XmaI和SmaI）克隆时，产生了四环素耐药转化子。
An Escherichia coli plasmid vector, pUN121, has been constructed which allows for positive selection of transformants harboring DNA inserts. The positive selection of transformants harboring DNA inserts. The vector is based on plasmid pTR262 (Roberts et al. Gene, 12, (1980), 123-127) in which the tetracycline resistance gene is under transcriptional control of the repressor protein coded by the phage lambda cI gene. This plasmid has been rearranged, using in vitro recombinant techniques including oligonucleotide mediated mutagenesis to yield a smaller plasmid (4.4 kb) with unique cloning sites for EcoRI, XmaI and SmaI in addition to the unique HindIII and BclI sites. The plasmid has a functional ampicillin resistance gene and the new restriction sites (EcoRI, XmaI and SmaI) when used for cloning, give rise to tetracycline resistant transformants.