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鼠伤寒沙门氏菌组氨酸操纵子第一部分的克隆与限制酶图谱

Cloning and restriction map of the first part of the histidine operon of Salmonella typhimurium.

作者信息

Barnes W M

出版信息

J Bacteriol. 1981 Jul;147(1):124-34. doi: 10.1128/jb.147.1.124-134.1981.

Abstract

The first part of the histidine operon of Salmonella typhimurium, hisGpeaGD, has been cloned onto the vector plasmid mini-ColE1 (pVH51). The resulting plasmid, pWB91, has a single EcoRI site and is 11,500 base pairs in size. The HindII restriction map was determined by the method of two-dimensional cross-annealing between a partial digest pattern and a complete digest pattern. The restriction fragment containing the genetic control region was identified with the aid of the small (35-base pair) internal deletion 01242 and the observation that heteroduplexed restriction fragments containing this deletion have markedly reduced mobility on polyacrylamide gels. The genetic control region was then mapped in more detail with other restriction enzymes. The genetic orientation of the restriction map was determined with the aid of several deletions of integral HindII fragments generated in vitro.

摘要

鼠伤寒沙门氏菌组氨酸操纵子的第一部分hisGpeaGD已被克隆到载体质粒mini-ColE1(pVH51)上。所得质粒pWB91有一个单一的EcoRI位点,大小为11,500碱基对。通过部分消化图谱和完全消化图谱之间的二维交叉退火方法确定了HindII限制性图谱。借助小的(35碱基对)内部缺失01242以及含有该缺失的异源双链限制性片段在聚丙烯酰胺凝胶上迁移率明显降低的观察结果,鉴定出了包含遗传控制区的限制性片段。然后用其他限制性酶对遗传控制区进行了更详细的图谱绘制。借助体外产生的几个完整HindII片段的缺失,确定了限制性图谱的遗传方向。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/e221/216016/bd1e6a598acb/jbacter00266-0135-a.jpg

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