大肠杆菌中苏氨酸脱氢酶结构基因的定位
Localization of the structural gene for threonine dehydrogenase in Escherichia coli.
作者信息
Ravnikar P D, Somerville R L
出版信息
J Bacteriol. 1986 Oct;168(1):434-6. doi: 10.1128/jb.168.1.434-436.1986.
Abstract
The threonine dehydrogenase (tdh) gene of Escherichia coli, cloned within the plasmid pDR121, was inactivated in vitro by inserting a segment of DNA carrying the chloramphenicol acetyltransferase (cat) gene. The insertionally inactivated tdh gene was then transferred by homologous recombination into the E. coli chromosome by the procedure of Winans et al. (J. Bacteriol. 161:1219-1221, 1985). Mating experiments, followed by P1-mediated two- and three-point crosses, enabled us to localize tdh near min 81.2. The order with respect to known markers is mtl-cysE-tdh-pyrE.
摘要
克隆于质粒pDR121中的大肠杆菌苏氨酸脱氢酶（tdh）基因，通过插入一段携带氯霉素乙酰转移酶（cat）基因的DNA在体外使其失活。然后，通过Winans等人（《细菌学杂志》161:1219 - 1221, 1985）的方法，将插入失活的tdh基因通过同源重组转移到大肠杆菌染色体中。通过接合实验，随后进行P1介导的两点和三点杂交，我们能够将tdh定位在约81.2分钟处。相对于已知标记的顺序是mtl - cysE - tdh - pyrE。
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