嗜热栖热放线菌中pyrE基因的分子克隆。
Molecular cloning of the pyrE gene from the extreme thermophile Thermus flavus.
作者信息
Vonstein V, Johnson S P, Yu H, Casadaban M J, Pagratis N C, Weber J M, Demirjian D C
机构信息
ThermoGen, Inc., Chicago, Illinois 60612, USA.
出版信息
J Bacteriol. 1995 Aug;177(15):4540-3. doi: 10.1128/jb.177.15.4540-4543.1995.
Abstract
Mutants of the extreme thermophile Thermus flavus in the pyrimidine biosynthetic pathway (Pyr-) were isolated by resistance to 5-fluoroorotic acid. The pyrE gene, which codes for the orotate phosphoribosyltransferase, was cloned by recombination with one of the isolated Pyr- T. flavus mutant strains. It was subcloned by complementation of an Escherichia coli pyrE mutant strain and was sequenced. The deduced polypeptide sequence extends over 183 amino acids. Several independent Pyr- mutations were mapped within the pyrE locus by recombination with fragments of the cloned gene.
摘要
通过对5-氟乳清酸的抗性筛选，分离出了嗜热栖热菌嘧啶生物合成途径中的突变体（Pyr-）。编码乳清酸磷酸核糖转移酶的pyrE基因，通过与一株分离得到的嗜热栖热菌Pyr-突变体菌株进行重组而被克隆。通过对大肠杆菌pyrE突变体菌株进行互补作用，将其亚克隆并测序。推导的多肽序列延伸超过183个氨基酸。通过与克隆基因的片段进行重组，在pyrE基因座内定位了几个独立的Pyr-突变。
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