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Related Concept Videos

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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.
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Groups of proteins may form a complex where each protein in this complex has a different role in the overall execution of the complex’s function. Often some of the proteins in the complex can be replaced by a closely related variant to give a complex that contains many of the same components yet is functionally distinct.
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A type of Lewis acid-base chemistry involves the formation of a complex ion (or a coordination complex) comprising a central atom, typically a transition metal cation, surrounded by ions or molecules called ligands. These ligands can be neutral molecules like H2O or NH3, or ions such as CN− or OH−. Often, the ligands act as Lewis bases, donating a pair of electrons to the central atom. These types of Lewis acid-base reactions are examples of a broad subdiscipline called coordination...
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Cruise control systems in cars are designed as multi-input systems to maintain a driver's desired speed while compensating for external disturbances such as changes in terrain. The block diagram for a cruise control system typically includes two main inputs: the desired speed set by the driver and any external disturbances, such as the incline of the road. By adjusting the engine throttle, the system maintains the vehicle's speed as close to the desired value as possible.
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Complex Numbers01:29

Complex Numbers

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The real number system cannot represent the square root of a negative number, which restricts solutions for certain equations, such as quadratics with negative discriminants. To address this, the complex number system was developed, introducing the imaginary unit i, where i = √(-1). This extension allows for the representation of all roots, including those involving negative radicands.A complex number is written in the form x + yi, where x and y are real numbers. Here, x represents the...
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What is Gene Expression?01:42

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Overview
Gene expression is the process in which DNA directs the synthesis of functional products, that is, proteins. Cells can regulate gene expression at various stages. It allows organisms to generate different cell types and enables cells to adapt to internal and external factors.
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Characterization of Multi-subunit Protein Complexes of Human MxA Using Non-denaturing Polyacrylamide Gel-electrophoresis
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Characterization of Multi-subunit Protein Complexes of Human MxA Using Non-denaturing Polyacrylamide Gel-electrophoresis

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Expressing Multi-subunit Complexes Using biGBac.

Florian Weissmann1, Jan-Michael Peters2

  • 1Research Institute of Molecular Pathology (IMP), Vienna Biocenter (VBC), Vienna, Austria.

Methods in Molecular Biology (Clifton, N.J.)
|April 2, 2018
PubMed
Summary
This summary is machine-generated.

The biGBac cloning method enables coexpression of up to 25 protein subunits from a single baculoviral transfer vector. This technique simplifies the production of complex protein assemblies using a hierarchical, two-step Gibson assembly process.

Keywords:
BEVSBaculovirus-insect cell expressionGibson assemblyMultigene expressionProtein complex

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Area of Science:

  • Molecular Biology
  • Protein Biochemistry
  • Gene Cloning

Background:

  • Recombinant protein complex reconstitution requires coexpression of multiple subunits.
  • Existing methods for coexpression can be complex and inefficient.

Purpose of the Study:

  • To describe a detailed protocol for the biGBac cloning method.
  • To enable efficient generation of baculoviral transfer vectors for complex protein assembly.

Main Methods:

  • The biGBac method utilizes Gibson assembly reactions and optimized DNA linkers.
  • It employs a hierarchical, two-step assembly strategy.
  • Up to five expression cassettes form a polygene cassette in step one; up to five polygene cassettes form the final vector in step two.

Main Results:

  • The biGBac method allows the generation of transfer vectors encoding up to 25 protein subunits.
  • This facilitates the coexpression of complex protein assemblies from a single vector.

Conclusions:

  • The biGBac cloning method provides an efficient and streamlined approach for producing recombinant protein complexes.
  • This protocol simplifies the generation of multi-subunit protein expression vectors.