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Assembly of Complex Microtubule Structures01:32

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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.
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Updated Protocol for the Assembly and Use of the Minibioreactor Array (MBRA)
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Mobius Assembly.

Andreas I Andreou1, Naomi Nakayama2,3

  • 1SyntheSys Centre for Synthetic and Systems Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, UK. Andreas.A@protonmail.com.

Methods in Molecular Biology (Clifton, N.J.)
|August 19, 2020
PubMed
Summary
This summary is machine-generated.

Mobius Assembly offers a user-friendly DNA assembly method for quickly creating DNA constructs. This system streamlines cloning with high capacity and modular design, simplifying complex genetic engineering.

Keywords:
AarIChromoproteinsDNA assemblyGolden Gate AssemblyMobius AssemblyMolecular cloningPhytobricksSynthetic biology

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Area of Science:

  • Synthetic Biology
  • Molecular Biology
  • Genetic Engineering

Background:

  • DNA construct generation is crucial for biological research and engineering.
  • Existing DNA assembly methods can be complex and time-consuming.
  • There is a need for efficient and user-friendly DNA assembly tools.

Purpose of the Study:

  • To introduce Mobius Assembly, a novel DNA assembly method.
  • To highlight the versatility and user-friendliness of Mobius Assembly.
  • To demonstrate its utility in streamlining DNA construct generation and combinatorial assemblies.

Main Methods:

  • Utilizes a two-level hierarchical modular cloning system.
  • Employs 4 bp standard overhangs defined by Phytobricks for part sharing.
  • Incorporates chromogenic proteins for positive assembly identification.
  • Designed for compatibility with different biological chassis.
  • Reduces domestication requirements for DNA parts.

Main Results:

  • Facilitates rapid and simple generation of DNA constructs.
  • Enables high cloning capacity and combinatorial assemblies.
  • Supports quadruple assembly augmentation.
  • Promotes standard part sharing through defined overhangs.
  • Simplifies identification of successful DNA assemblies.

Conclusions:

  • Mobius Assembly is a versatile and efficient method for DNA construct generation.
  • The system simplifies complex cloning procedures, making it accessible for various applications.
  • It offers advantages in speed, capacity, and ease of use compared to traditional methods.