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氯霉素乙酰转移酶基因在胞嘧啶取代的T4噬菌体中的克隆与表达。

Cloning and expression of a chloramphenicol acetyltransferase gene in cytosine-substituted T4 bacteriophage.

作者信息

Noguchi T, Takahashi H, Saito H

出版信息

Gene. 1986;44(1):133-8. doi: 10.1016/0378-1119(86)90052-1.

Abstract

We have developed an efficient method for transferring foreign genes into the T4 phage genome. Any foreign genes inserted into the T4 uvsY gene cloned on plasmids can be transferred into a cytosine-substituted T4dC(delta NB5060) phage genome by a replacement type of recombination. To achieve this, we constructed chimeric plasmids which had a chloramphenicol acetyltransferase gene (cat) derived from transposon Tn9 inserted into the Bg/II site within the T4 uvsY gene on pBR322. The cat gene was then transferred by in vivo recombination into the T4dC(delta NB5060) phage genome. Moreover, it was demonstrated that the cat gene in the hybrid T4dC phage was expressed upon phage infection and development.

摘要

我们已经开发出一种将外源基因导入T4噬菌体基因组的有效方法。任何插入到克隆在质粒上的T4 uvsY基因中的外源基因,都可以通过替换型重组转移到胞嘧啶取代的T4dC(δNB5060)噬菌体基因组中。为了实现这一点,我们构建了嵌合质粒,该质粒具有源自转座子Tn9的氯霉素乙酰转移酶基因(cat),插入到pBR322上T4 uvsY基因内的Bg/II位点。然后,cat基因通过体内重组转移到T4dC(δNB5060)噬菌体基因组中。此外,还证明了杂交T4dC噬菌体中的cat基因在噬菌体感染和发育时表达。

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