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相关概念视频

RNA-seq03:21

RNA-seq

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RNA sequencing, or RNA-Seq, is a high-throughput sequencing technology used to study the transcriptome of a cell. Transcriptomics helps to interpret the functional elements of a genome and identify the molecular constituents of an organism. Additionally, it also helps in understanding the development of an organism and the occurrence of diseases. 
Before the discovery of RNA-seq, microarray-based methods and Sanger sequencing were used for transcriptome analysis. However, while...
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相关实验视频

Updated: Jul 24, 2025

Nanopore DNA Sequencing for Metagenomic Soil Analysis
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为蛋白质工程应用开发一个纳米孔测序工作流程

Daniel Zhi Jun Tan1, Vincent Fung1, Tao Sun1

  • 1Department of Chemical and Biomolecular Engineering, National University of Singapore, Singapore 117585, Singapore.

ACS synthetic biology
|July 5, 2023
PubMed
概括
此摘要是机器生成的。

与Illumina测序相比,纳米孔测序在蛋白质工程突变库中显著减少了链交换错误. 一个优化的纳米孔工作流程有助于识别高度活跃的酒精脱酶突变体.

关键词:
埃舍里希亚大肠杆菌 (Escherichia coli) 是一个大肠杆菌.基于增长的选择.纳米孔测序的测序蛋白质工程工程 蛋白质工程二次性酒精脱酶的使用位点和突变发生的突变发生.

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科学领域:

  • 生物技术是生物技术.
  • 分子生物学分子生物学
  • 蛋白质工程是指蛋白质工程.

背景情况:

  • 下一代测序 (NGS) 对于识别蛋白质工程中的遗传突变至关重要.
  • 商业上可用的NGS技术如Illumina和纳米孔测序具有不同的错误概况.
  • 字符串交换是测序突变库的一个已知的问题,可能会混结果.

研究的目的:

  • 评估和比较Illumina和纳米孔测序的性能,用于分析蛋白质工程突变库.
  • 开发和优化用于纳米孔测序的库准备工作流程,以最大限度地减少链交换.
  • 为了证明优化的纳米孔工作流在识别改进的蛋白质变体中的实用性.

主要方法:

  • 在现有和内部构建的突变库上对Illumina NGS和纳米孔测序进行比较分析.
  • 开发一种用于纳米孔测序的新型库准备工作流程.
  • 应用优化的工作流程,以增长为基础的酒精脱酶突变体的选择.
  • 使用纳米孔测序数据量化突变丰富折叠变化.

主要成果:

  • 照明测序显示了大量的读数与链交换,混合突变信息.
  • 与Illumina相比,纳米孔测序证明了链交换的显著减少.
  • 优化的纳米孔库准备工作流进一步降低了链交换发生率.
  • 工作流成功量化了1728个突变物在选择过程中的丰富折叠变化.
  • 使用折叠变化数据确定了一种高度活跃的酒精脱酶突变体 (>500%的改善).

结论:

  • 纳米孔测序提供了一种更准确的方法来分析蛋白质工程突变库,因为减少了链交换.
  • 开发的纳米孔库准备工作流程提高了蛋白质工程的准确性和效率.
  • 这种快速而实惠的测序方法对于选择改进的蛋白质变体是有价值的,特别是当活性与生长相关时.