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相关概念视频

Oligosaccharide Assembly01:24

Oligosaccharide Assembly

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Protein glycosylation starts in the ER lumen and continues in the Golgi apparatus. Glycosyltransferases catalyze the addition of sugar molecules or glycosylation of proteins. Usually, these enzymes add sugars to the hydroxyl groups of selected serine or threonine residues to form O-linked glycans or the amino groups of asparagine residues to form N-linked glycans. Different positions on the same polypeptide chain can contain differently linked glycans.
Multiple sugar molecules that may or may...
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Protein Complex Assembly02:41

Protein Complex Assembly

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Proteins can form homomeric complexes with another unit of the same protein or heteromeric complexes with different types.  Most protein complexes self-assemble spontaneously via ordered pathways, while some proteins need assembly factors that guide their proper assembly. Despite the crowded intracellular environment, proteins usually interact with their correct partners and form functional complexes.
Many viruses self-assemble into a fully functional unit using the infected host cell to...
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相关实验视频

Updated: Jul 15, 2025

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning
08:31

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

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高复杂度的一金门大门装配组件

Andrew P Sikkema1, S Kasra Tabatabaei1, Yan-Jiun Lee1

  • 1Research Department, New England Biolabs, Ipswich, Massachusetts, USA.

Current protocols
|September 27, 2023
PubMed
概括
此摘要是机器生成的。

这项研究提高了DNA组装使用数据优化设计 (DAD) 在复杂的金门组装项目高精度. DAD能够有效地连接许多DNA片段,提高克隆产量和成功率.

关键词:
黄金大门大会的会议会议.数据优化的组装设计设计.多基因结构构建了多基因结构.小基因组组装小基因组组合成生物学 合成生物学

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Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit
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Mouse Genome Engineering Using Designer Nucleases
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Mouse Genome Engineering Using Designer Nucleases

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相关实验视频

Last Updated: Jul 15, 2025

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Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

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Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit
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Genetic Modification of Cyanobacteria by Conjugation Using the CyanoGate Modular Cloning Toolkit

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Mouse Genome Engineering Using Designer Nucleases
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Mouse Genome Engineering Using Designer Nucleases

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科学领域:

  • 分子生物学分子生物学
  • 合成生物学 合成生物学
  • 生物技术是生物技术.

背景情况:

  • 金门组合是一种多功能DNA克隆方法,使用IIS型限制酶在单一反应中连接多个DNA片段.
  • 传统的金门组装方法在组装大量碎片 (通常为5-8个) 时,在准确性和产量方面面临限制.
  • 最近的进展表明,有可能进行高度复杂的组装,以更高的精度连接多达52个碎片.

研究的目的:

  • 描述应用数据优化装配设计 (DAD) 原则和在线工具以增强金门装配的方法.
  • 为了评估和优化融合站点集,划分基因组序列,并为复杂的DNA结构设计一组件.
  • 为高复杂性组装提供协议,包括T7菌体基因组组装,以及评估成功的方法.

主要方法:

  • 应用DAD原则和在线工具 (NEBridge Ligase Fidelity Viewer,GetSet工具,SplitSet工具) 来评估和选择聚变地点.
  • 设计和执行单金门组件的中高复杂度 (12-36片段),包括小基因组组件.
  • 用于生成高纯度DNA部分,量化DNA,可视化组件和使用长读序列测序验证结果的协议.

主要成果:

  • 通过使用DAD连接多达52个碎片,证明了复杂组件的高组装精度.
  • 开发并介绍了优化融合站点,分割基因组序列和执行单组件的方法.
  • 从多个部分成功组装了T7菌体基因组,展示了DAD原则的实际应用.

结论:

  • 数据优化装配设计 (DAD) 显著提高了复杂的金门装配的准确性和效率.
  • 描述的方法和在线工具有助于设计和执行具有挑战性的DNA组装项目.
  • 这种方法可以高准确地构建复杂的DNA分子,包括整个基因组.