Prentki P, Krisch H M
Gene. 1982 Feb;17(2):189-96. doi: 10.1016/0378-1119(82)90072-5.
The construction of a plasmid vector which facilitates the cloning and recovery of blunt-ended DNA fragments is described. This plasmid, called pHP34, differs from pBR322 by a 10-bp insertion which introduces a unique SmaI site immediately flanked by two EcoRI sites. Blunt-ended DNA fragments cloned in the SmaI site can be recovered by digestion with EcoRI. Small cloned fragments can be chemically sequenced using a strategy which does not require their purification. The use of a plasmid related to pHP34 for in vitro mutagenesis by the insertion of a DNA linker fragment conferring an antibiotic resistance is also discussed.
本文描述了一种有助于克隆和平端DNA片段回收的质粒载体的构建。这种名为pHP34的质粒与pBR322的区别在于有一个10碱基对的插入片段，该片段引入了一个独特的SmaI位点，其两侧紧邻两个EcoRI位点。克隆到SmaI位点的平端DNA片段可通过用EcoRI酶切回收。小的克隆片段可用一种无需纯化的策略进行化学测序。还讨论了使用与pHP34相关的质粒通过插入赋予抗生素抗性的DNA接头片段进行体外诱变的方法。
