Recombinant Taq DNA Polymerase TaKaRa Taq^TM

Company: Takara Bio

Catalog#: R001WZ

In vitro Glutamylation Inhibition of Ubiquitin Modification and Phosphoribosyl-Ubiquitin Ligation Mediated by Legionella pneumophila Effectors

Alan G. Sulpizio, Marena E. Minelli and Yuxin Mao

Published online: 2020-11-05

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Characterization of Protein Domain Function via in vitro DNA Shuffling

Author:

Kathy Hiu Laam Po, Edward Wai Chi Chan and Sheng Chen,

Date:

2018-06-05

[Abstract] We recently investigated the molecular events that drive evolution of the CTX-M-type β-lactamases by DNA shuffling of fragments of the bla_CTX-M-14 and bla_CTX-M-15 genes. Analysis of a total of 51 hybrid enzymes showed that enzymatic activity could be maintained in most cases, yet the enzymatically active hybrids were found to possess much fewer amino acid substitutions than the few hybrids that became inactive, suggesting that point mutations in the constructs rather than reshuffling of the fragments of the two target genes would more likely cause disruption of CTX-M activity. Certain important residues that played important functional roles in mediating enzyme activity were identified. These findings suggest that DNA shuffling is an effective ... [摘要] 我们最近研究了通过对CTX-M-14和EMX-M-14的片段进行DNA改组来驱动CTX-M型β-内酰胺酶进化的分子事件， bla CTX-M-15基因。总共51种杂合酶的分析显示酶活性在大多数情况下可以保持，但是酶活性杂合体被发现比少数杂交体具有少得多的氨基酸取代，这表明构建体中的点突变而不是两个靶基因片段的重新洗牌将更可能导致CTX-M活性的破坏。确定了一些在介导酶活性中起重要作用的重要残基。这些发现表明，DNA改组是一种有效的方法来鉴定和表征细菌蛋白质中的重要功能结构域。

【背景】DNA重组是一种自然过程，通过该过程，细菌之间交换遗传物质以增强环境压力下的生存适应性。几种杂交CTX-M-内酰胺酶（CTX-M-64，CTX-M-123，CTX-M-137和CTX-M-132）可能是由bla CTX-M-14和 bla CTX-M-15基因是世界上最常见的变异体，近年来已有报道（Nagano et al。，2009; Tian et al。，2014; He et al。，2015; Liu et。， 2015年）。在这些杂合酶中，包含CTX-M-15的N-和C-末端部分和CTX-M-14的中间片段的CTX-M-64显示出比其亲本原型更高的催化活性（He <等）。，2015）。

DNA改组是一种分子途径，被设计为通过PCR介导的两种靶基因的随机组合来模拟和加速进化过程（Crameri ...

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Recombinant Taq DNA Polymerase TaKaRa TaqTM

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Recombinant Taq DNA Polymerase TaKaRa Taq^TM