Marsh J L, Erfle M, Wykes E J
Gene. 1984 Dec;32(3):481-5. doi: 10.1016/0378-1119(84)90022-2.
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.
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）。
