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利用确定基因末端的缺失对大肠杆菌gnd基因中的插入突变进行定位。

Mapping of insertion mutations in gnd of Escherichia coli with deletions defining the ends of the gene.

作者信息

Wolf R E, Cool J A

出版信息

J Bacteriol. 1980 Mar;141(3):1222-9. doi: 10.1128/jb.141.3.1222-1229.1980.

Abstract

A genetic map was prepared for gnd, the gene of Escherichia coli which encodes the metabolically regulated 6-phosphogluconate dehydrogenase. Direct selection methods were used for the isolation of mutants with deletions that define the respective ends of gnd. These selections depended on the availability of a defective lysogen in which gnd was present on a lambda h80 dgnd his prophage located at the att phi 80 region of the chromosome. Mutants with deletions entering gnd from the his-distal end were selected as Gnd- TonB- mutants. Mutants with his-proximal gnd deletions were selected as Gnd-, temperature-resistant mutants of a specially prepared stable lysogen. Gnd- mutants were also isolated after mutagenesis with bacteriophage Mu cts61, and genetic tests were used to determine which mutants carry a Mu cts61 prophage in gnd. The deletion mutations were mapped against each other and against the insertion mutations through the use of F' merodiploid strains. The insertion mutations mapped at seven distinct sites in gnd; three mapped under the deletions defining the his-proximal portion of the gene and three mapped with the his-distal deletions.

摘要

为大肠杆菌中编码代谢调控的6-磷酸葡萄糖酸脱氢酶的gnd基因构建了遗传图谱。采用直接选择法分离出具有缺失的突变体,这些缺失确定了gnd基因的各个末端。这些选择依赖于一种缺陷溶原菌的存在,其中gnd基因位于位于染色体att phi 80区域的λh80 dgnd his原噬菌体上。从his远端进入gnd的缺失突变体被选为Gnd-TonB-突变体。his近端gnd缺失的突变体被选为Gnd-,即一种特别制备的稳定溶原菌的温度抗性突变体。在用噬菌体Mu cts61诱变后也分离出了Gnd-突变体,并通过遗传测试确定哪些突变体在gnd中携带Mu cts61原噬菌体。通过使用F'部分二倍体菌株,将缺失突变相互之间以及与插入突变进行了定位。插入突变定位在gnd的七个不同位点;三个位于定义基因his近端部分的缺失之下,三个与his远端缺失定位在一起。

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