Changes in the substrate specificities of an enzyme during directed evolution of new functions.

Changes in the substrate specificities of an enzyme during directed evolution of new functions.
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新功能定向进化过程中酶底物特异性的变化。

DOI:
10.1021/bi00517a015
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发表时间:
1981
期刊:
影响因子:
2.9
通讯作者:
Hall,BG
Hall,BG
中科院分区:
生物学3区
文献类型:
--
作者:
Hall,BG

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大肠杆菌K12的ebg系统被用作研究新代谢功能的获得性进化的模型。从E.在携带lacZ(β-半乳糖苷酶)基因内的大缺失的大肠杆菌K12中,我选择了一系列自发突变菌株,这些菌株已经进化出利用乳糖和其它β-半乳糖苷糖作为唯一碳源和能源的能力(Hall & Hard,1974; Hall,1976 a,B,1977,1978; Hall & Zuzel,1980)。自发突变发生在位于E. coli图谱(Hall & Hard,1975; Bachmann & Low,1980)。操纵子受紧密连锁的ebgR基因产物的负控制(Hall & Hard,1975)。ebgR突变对进化过程很重要(Hall & Clarke,1977),但不在本文讨论范围内。感兴趣的结构基因是ebgA,它指定了120000分子量的多肽,该多肽是同六聚体ebg/β-半乳糖苷酶的亚基(Hall,f,来自微生物学部分U-44,生物科学组,
The ebg system of Escherichia coli K12 is being used as a model to study the acquisitive evolution of new metabolic functions. Beginning with a strain of E. coli K12 bearing a large deletion within the lacZ (/3-galactosidase) gene, I have selected a series of spontaneous mutant strains that have evolved the ability to utilize lactose and other/3-galactoside sugars as sole carbon and energy sources (Hall & Hard, 1974; Hall, 1976a, b, 1977, 1978; Hall & Zuzel, 1980). The spontaneous mutations occur in theebg (evolved/3-galactosidase) operon located at 66 min on the E. coli map (Hall & Hard, 1975; Bachmann & Low, 1980). The operon is under negative control by the product of the tightly linked ebgR gene (Hall & Hard, 1975). Mutations in ebgR are important to the evolutionary process (Hall & Clarke, 1977) but are outside the scope of this paper. The structural gene of interest is ebgA which specifies a 120000 molecular weight polypeptide that is the subunit of the homohexameric ebg/3-galactosidase (Hall, f From the Microbiology Section U-44, Biological Sciences Group,
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