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具有多用途克隆位点的pIC质粒和噬菌体载体,用于通过插入失活进行重组体筛选。

The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation.

作者信息

Marsh J L, Erfle M, Wykes E J

出版信息

Gene. 1984 Dec;32(3):481-5. doi: 10.1016/0378-1119(84)90022-2.

Abstract

The versatility of insertional inactivation of beta-galactosidase activity for subcloning and sequencing has been enhanced by combining a chemically synthesized oligonucleotide which specifies nine 6-bp-cutter restriction sites including BglII, XhoI, NruI, ClaI, SacI and EcoRV in various configurations with existing polylinkers to create a set of highly versatile cloning sites. These improved polylinkers have been inserted into plasmids (the pICs) for routine cloning of double-stranded DNA, and into chimeric phage/plasmids (the pICEMs) for biological production of single stranded DNA. The most versatile polylinker specifies 17 restriction sites in the beta-galactosidase alpha-complementing gene fragment. One of the new polylinkers was inserted into M13 DNA to produce a vector (M13mIC7) with nine cloning sites.

摘要

通过将化学合成的寡核苷酸与现有的多克隆位点相结合,β-半乳糖苷酶活性插入失活在亚克隆和测序中的通用性得到了增强。该寡核苷酸指定了九个6碱基切割限制位点,包括BglII、XhoI、NruI、ClaI、SacI和EcoRV,它们以各种构型排列,从而创建了一组高度通用的克隆位点。这些改进的多克隆位点已被插入质粒(pICs)用于双链DNA的常规克隆,并被插入嵌合噬菌体/质粒(pICEMs)用于单链DNA的生物制备。最通用的多克隆位点在β-半乳糖苷酶α-互补基因片段中指定了17个限制位点。其中一个新的多克隆位点被插入到M13 DNA中,以产生一个具有九个克隆位点的载体(M13mIC7)。

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