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噬菌粒:柯斯质粒(ColE1质粒)与大肠杆菌噬菌体λ之间的杂种。

Phasmids: hybrids between ColE1 plasmids and E. coli bacteriophage lambda.

作者信息

Brenner S, Cesareni G, Karn J

出版信息

Gene. 1982 Jan;17(1):27-44. doi: 10.1016/0378-1119(82)90098-1.

Abstract

Plasmids carrying cloned lambda att sites may be integrated into the bacteriophage genome by the site-specific recombination mechanism of lambda. The cross, referred to as "lifting" the plasmid, requires mixed infection of an Escherichia coli strain carrying the plasmid with two appropriately constructed "lifting" lambda phages. One phage donates a short left arm and the other donates a short right arm. These two short arms are of insufficient length to produce a viable phage genome and yield no recombinants when crossed on standard bacteria. However, viable recombinants are obtained when the genome length is extended by integration of one or more plasmids. We call these recombinants phasmids. They contain multiple att sites introduced at the ends of the integrated plasmids, and in the presence of integrase, recombination between these att sites can be exploited to effect release of the plasmid components. These novel genetic elements can be used in a variety of ways as vectors in genetic manipulation experiments. Sequences cloned in phasmids may be studied as a component of either a plasmid and or of a phage, and easily interconverted between the two states.

摘要

携带克隆的λ附着位点的质粒可通过λ的位点特异性重组机制整合到噬菌体基因组中。这种将质粒“提升”的杂交过程需要用两种经过适当构建的“提升”λ噬菌体对携带该质粒的大肠杆菌菌株进行混合感染。一种噬菌体提供一个短的左臂,另一种提供一个短的右臂。这两个短臂的长度不足以产生一个有活力的噬菌体基因组,并且在标准细菌上杂交时不会产生重组体。然而,当通过整合一个或多个质粒来延长基因组长度时,可获得有活力的重组体。我们将这些重组体称为噬菌粒。它们含有在整合质粒末端引入的多个附着位点,并且在整合酶存在的情况下,可利用这些附着位点之间的重组来实现质粒组分的释放。这些新型遗传元件在基因操作实验中可用作多种载体。克隆在噬菌粒中的序列可作为质粒或噬菌体的一个组分进行研究,并且可以很容易地在这两种状态之间相互转换。

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