Directed evolution of cellobiose utilization in Escherichia coli K12.

Directed evolution of cellobiose utilization in Escherichia coli K12.
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大肠杆菌 K12 中纤维二糖利用的定向进化。

DOI:
--
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发表时间:
1984
影响因子:
10.7
通讯作者:
B. Hall
B. Hall
中科院分区:
生物学1区
文献类型:
--
作者:
M. Kricker;B. Hall

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大肠杆菌K12的纤维二糖分解代谢系统正被用于研究隐藏基因在新功能进化中的作用。大肠杆菌不使用β-葡萄糖苷糖;然而,几个位点的突变可以激活隐蔽的bgl操纵子,并允许在β-葡萄糖苷糖熊果苷和水杨苷上生长。这样的Bgl+突变体不使用纤维二糖,纤维二糖是自然界中最常见的β-葡糖苷。我们从大肠杆菌Bgl ~+突变株中分离到一株Cel ~+(利用纤维二糖)突变株。coli K12中表达。Cel+突变体在纤维二糖、熊果苷和水杨苷上生长良好。利用这些β-葡萄糖苷的基因位于E. coli作图。bgl操纵子的基因不参与纤维二糖的利用。引入覆盖bgl的缺失不影响利用纤维二糖、熊果苷或水杨苷的能力,表明新的Cel+基因提供所有三种功能。自发的纤维二糖阴性突变体也变成熊果苷和水杨苷阴性。对这些突变体的β-葡糖苷阳性回复突变体的分析表明,存在用于利用每种β-葡糖苷糖的单独的基因座。这些基因紧密相连,可以从一个位点激活。在未知位置的第四个基因增加了纤维二糖的生长速率。cel基因构成了E. coli K12。
The cellobiose catabolic system of Escherichia coli K12 is being used to study the role of cryptic genes in evolution of new functions. Escherichia coli does not use beta-glucoside sugars; however, mutations in several loci can activate the cryptic bgl operon and permit growth on the beta-glucoside sugars arbutin and salicin. Such Bgl+ mutants do not use cellobiose, which is the most common beta-glucoside in nature. We have isolated a Cel+ (cellobiose-utilizing) mutant from a Bgl+ mutant of E. coli K12. The Cel+ mutant grows well on cellobiose, arbutin, and salicin. Genes for utilization of these beta-glucosides are located at 37.8 min on the E. coli map. The genes of the bgl operon are not involved in cellobiose utilization. Introduction of a deletion covering bgl does not affect the ability to utilize cellobiose, arbutin, or salicin, indicating that the new Cel+ genes provide all three functions. Spontaneous cellobiose negative mutants also become arbutin and salicin negative. Analysis of beta-glucoside positive revertants of these mutants indicates that there are separate loci for utilization of each of the beta-glucoside sugars. The genes are closely linked and may be activated from a single locus. A fourth gene at an unknown location increases the growth rate on cellobiose. The cel genes constitute a second cryptic system for beta-glucoside utilization in E. coli K12.
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DOI: 10.1099/00221287-121-1-85
发表时间: 1980
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影响因子: --
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DOI: 10.1093/oxfordjournals.molbev.a040300
发表时间: 1983
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