Applied Biochemistry at Sheffield Hallam University | Flashcards & Summaries

Applied Biochemistry at Sheffield Hallam University

Flashcards and summaries for Applied Biochemistry at the Sheffield Hallam University

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Exemplary flashcards for Applied Biochemistry at the Sheffield Hallam University on StudySmarter:

What are the four steps of translation? 

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How does eIF2α kinase upregulate mRNA-specific translation?

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Describe Lipofection
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Exemplary flashcards for Applied Biochemistry at the Sheffield Hallam University on StudySmarter:

What is bactofection? How does it work?

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What are the five steps of translational initiation?

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What is the concentration of ternary complex determined by?

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What is the disadvantage of microinjection?
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Exemplary flashcards for Applied Biochemistry at the Sheffield Hallam University on StudySmarter:

How is it possible to post-translationally modify recombinant proteins in bacteria?

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When using one plasmid to express two GOIs, what does the plasmid need two of?

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How does electroporation allow for gene transfer?

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What is the limiting factor in translational initiation? 
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Proteins need to be ____ and ____ to produce well-refracting crystals/

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Exemplary flashcards for Applied Biochemistry at the Sheffield Hallam University on StudySmarter:

Applied Biochemistry

What are the four steps of translation? 
  • Initiation
  • Elongation 
  • Termination
  • Ribosome recycling 

Applied Biochemistry

How does eIF2α kinase upregulate mRNA-specific translation?

  • eIF2α kinase phosphorylates eIF2α into eIF2(αP)
  • eIF2(αP) inhibits eIF2B, thus reducing the amount of ternary complex
  • This alters the translation of uORFs, leading to the up-expression of ATF4, which is a transcription activator

Applied Biochemistry

Describe Lipofection
The plasmid DNA is covered with lipids, allowing entrance into the cell.

Applied Biochemistry

What is bactofection? How does it work?
The use of bacteria that invade human cells to transfer genes. The bacteria enters the cell and undergoes lysis with the cell, releasing the plasmid.

Applied Biochemistry

What are the five steps of translational initiation?

  1. Ternary complex formation
    • Consists of: eIF2, Met-tRNA, and GTP
    • Met-tRNA is delivered by eIF2
  2. 43S preinitiation complex formation
    • Met-tRNA is delivered by eIF2-GTP to the ribosomal subunit 40S
  3. Recruitment of 43S complex to mRNA
    • at the 5' end
    • by cap-binding proteins (i.e. eIF3, eIF4)
  4. Scanning and AUG recognition
    • scanning the 5' untranslated region
  5. Assembly of the 80S ribosome
    • the 60S ribosomal subunit joins

Applied Biochemistry

What is the concentration of ternary complex determined by?

eIF2B

Applied Biochemistry

What is the disadvantage of microinjection?
It can only be used on a few cells at a time.

Applied Biochemistry

How is it possible to post-translationally modify recombinant proteins in bacteria?
  • Added functionality is possible by co-expression
  • Co-express with a complex that modifies proteins post-translationally

Applied Biochemistry

When using one plasmid to express two GOIs, what does the plasmid need two of?
  • Two multiple cloning sites
  • Two different promoters

Applied Biochemistry

How does electroporation allow for gene transfer?
Creates of nanopores in plasma membrane by electrical pulses. These nanopores allow the DNA into the cell.

Applied Biochemistry

What is the limiting factor in translational initiation? 
Levels of ternary complex

Applied Biochemistry

Proteins need to be ____ and ____ to produce well-refracting crystals/

Pure and homogenous

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