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From Molecules to Ecosystems (Bio1007) > L. 22 - Cell Factories & Biotechnology > Flashcards

L. 22 - Cell Factories & Biotechnology Flashcards

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Organic Chemistry 101 flashcards
1
Q

L.O.

A
  • Explain why microbes are useful for biotechnology, using examples of specific fungi (cerevisiae) and bacteria (E.coli)
  • Explain what a plasmid is, and define the roles of different kinds of plasmids in nature and in biotechnology
  • Define the terms “DNA cloning”, “recombinant DNA”, “GMO”
  • Explain how recombinant DNA and GMOs are made, and especially which enzymes do which jobs in this process
  • Discuss why vaccines are important, and how recombinant DNA methods can be used to make them
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2
Q

Fermentation (traditional biochemistry)

A
  • Fermented alcohol and to preserve food
  • Mixed cultures of naturally occuring bacteria
  • Yeast fermentation is ancient ~7000BC China
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3
Q

Types of Biotechnology

A

Molecular biotechnology:
- Cellular machinery inside cells

Cellular biotechnology:
- Whole cells and microbes

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4
Q

Cellular biotechnology

A
  • Early/ ancient methods
  • Fermentation
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5
Q

Molecular biotechnology

A
  • Modern methods
  • Looking at manipulating microbes and enzymes
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6
Q

Microbes for biotechnology

A
  • Virus
  • Archaea
  • Algae
  • Fungi
  • Protists
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7
Q

Viruses for biotechnology

A

Viruses are used as vectors
Vectors = carry gene in DNA
- Source of enzymes (T4 Liagse, for ligating)

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8
Q

Archaea for biotechnology

A
  • Diverse living conditions
  • Source of enzymes (PCR, thermostable polymerase, to copy DNA)
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9
Q

Bacteria for biotechnology

A
  • Host for DNA cloning
  • Express proteins
  • Quick and easy to work with
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10
Q

Algae for biotechnology

A
  • Eukaryotes
  • Green biotechnology
  • Biofuels, (convert CO2 and light into biofuels)
  • Carbon negative
  • Low maintenance
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11
Q

Fungi for biotechnology

A

Yeast = Cloning and expression hosts, similar to bacteria
Moulds = antibiotic synthesis, penecillin

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12
Q

Host cell examples

A

Bacteria: (e-coli)
- Fast growing
- Easy to extract/ add plasmid DNA
- Model organisms

Yeasts: cerevisiae
- Bread and beer
- Express Eukayrotic genes (body recognise as eukaryotic)
- Generally safely recognised (food grade)

Biosynthesis of high-value products from simple raw materials. All starting from DNA

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13
Q

Vectors

A
  • Deliver DNA (usually foreign)

Plasmids:
- Circular DNA
- Replicate independently of chromosomes
- Most common vector

Viruses:
- ‘hijacking’ host cells

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14
Q

Plasmids

A

Naturally found and allow genes to be swapped (Horizontal gene transfer)

Non-essential genes for growth and division of bacteria (can help in certain conditions - antibiotic resistance)

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15
Q

Key factors of plasmids

A

Selectable (gene) marker:
- Forces bacteria to takeup and sue the plasmid for anitibiotic resistance/ metabolic activity

Replication function:
- Persistance in host
- Help bacteria keep the plasmid

Cloning site:
- Manipulate site
- Insert foreign genes

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16
Q

Molecular cloning

A
  • Make many copies of entity

3 Steps:
1. Recombinant DNA preperation (to be modified)
2. Transformation and screening
3. Copying and expression

17
Q

Molecular cloning step 1:

A

Recombinant DNA preperation

Recombinant DNA:
- Foreign DNA and plasmid vector mixed

Identify gene of interest:
- Digest DNA (cut with enzyme)
- Ligate DNA (join foreign DNA with vector)

18
Q

Molecular cloning step 2:

A

Transformation and screening:

Transformation:
- Host cell picks up recombinant DNA under stressful conditions

Screening:
- Selectrable (gene) marker to see it
- Transformation is NOT 100% efficient

eg.
antibiotic resistance
metabolic activity
PCR (thermostable polymerase)

19
Q

Molecular cloning step 3:

A

Copying and expressing:

Copies of organisms/ DNA:
- express molecules/ proteins

Cloning Site:
- induce growth/ gene expression
- lacZα gene

PCR:
- copying DNA

Express foreign protein:
- End product
- Used to make another product

20
Q

Molecular cloning end result

A

Genetically modified Organism (GMO):

  • Foreign gene inside

Risks:
- depends on gene added
- antibiotic resistance gene transfer into pathogens
- commercial risks

21
Q

Vaccines

A
  • Recombinant products
  • Some disease have no cure, prevention is only option
  • Vaccines lead to ‘herd immunity’
  • Adaptive immune response to recognise antigens associated with the pathogen
  • live or dead microbes
22
Q

Smallpox

A
  • Variola virus
  • Smallpox is erradicated due to vaccines
23
Q

Preparing a recombinant vaccine

A

1. Recombinant DNA preperation:
- identify gene of interest
- Digest DNA
- Ligate DNA

2. Transformation and screening:
- microbe takes up recombinant DNA
- select those that did

3. Copying and expressing:
- Express foreign gene
- purification
- Antigen product

[heft]
(O = Plasmid) .(≠ = DNA)
O + ≠≠≠≠

C + .≠ ≠ ≠ ≠ .<=Digest

C≠ .<=Ligate

From Molecules to Ecosystems (Bio1007) (25 decks)

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