Nieradko J, Podgórska B
Department of Microbiology, University of Gdańsk, Poland.
Acta Biochim Pol. 1993;40(2):273-8.
A fragment of T4 DNA (XbaI-HindIII) comprising the genes 51, 27, 28, which encodes the central plug proteins was cloned into plasmid pT7-5 and p7-6 (T7 RNA polymerase expressing system). The examined genes were only overexpressed when the orientation of cloned DNA to promoter phi 10 was as follows: promoter phi 10 and genes 51, 27, 28. This was achieved when the fragment (XbaI-HindIII) was cloned into plasmid pT7-5. Gene 27 and 28 were overexpressed when the intact fragment (XbaI-HindIII) was used. The high rate of the synthesis of proteins 27 and/or 28 had a strong inhibitory effect on the level of synthesis of the product of gene 51. For the overexpression of gene 51 in this system a deletion derivate which was devoid of gene 28 and a larger fragment of gene 27 was prepared.
包含编码中心塞蛋白的基因51、27、28的一段T4 DNA（XbaI-HindIII）被克隆到质粒pT7-5和p7-6（T7 RNA聚合酶表达系统）中。只有当克隆的DNA与启动子phi 10的方向如下时，所检测的基因才会过度表达：启动子phi 10以及基因51、27、28。当将片段（XbaI-HindIII）克隆到质粒pT7-5中时实现了这一点。当使用完整片段（XbaI-HindIII）时，基因27和28会过度表达。蛋白质27和/或28的高合成率对基因51产物的合成水平有强烈的抑制作用。为了在该系统中使基因51过度表达，制备了一个缺失基因28且包含更大片段基因27的缺失衍生物。
