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STB1073 Biotechnology: Gene Cloning
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STB1073 Biotechnology: Gene Cloning

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Questions and Answers

What is the primary function of Agrobacterium in biotechnology?

  • To produce antibiotics
  • To transfer genes to plants for genetic engineering (correct)
  • To produce industrial enzymes like amylase
  • To combat vitamin A deficiency in humans
  • Which of the following is NOT a contribution of microbiology to biotechnology?

  • Supply of antibiotics
  • Cloning process
  • Industrial enzymes production
  • Fuel production (correct)
  • What is the name of the disease caused by Agrobacterium in plants?

  • Tumour inducing disease
  • Vitamin A deficiency
  • Golden rice disease
  • Crown gall disease (correct)
  • What is the role of T-DNA in Agrobacterium?

    <p>To transfer genes to plants</p> Signup and view all the answers

    What is the purpose of 'disarming' the Ti plasmid?

    <p>To delete the tumour-inducing genes</p> Signup and view all the answers

    What is the size of the circular DNA plasmid responsible for tumour production and gene transfer?

    <p>200 kb</p> Signup and view all the answers

    Which of the following viruses is used as a vector in gene therapy?

    <p>Retroviruses</p> Signup and view all the answers

    What is the significance of Golden rice?

    <p>It is used to combat vitamin A deficiency</p> Signup and view all the answers

    What is the role of border repeats in T-DNA?

    <p>To facilitate plant transformation</p> Signup and view all the answers

    Which of the following is an application of microbiology in agriculture?

    <p>Genetic engineering for plant improvement</p> Signup and view all the answers

    Study Notes

    Modern Biotechnology: Multidisciplinary Input

    • Modern biotechnology involves the integration of biochemistry, genetics, microbiology, and cell biology.

    General Strategy in Biotechnology

    • The general strategy in biotechnology involves cloning a gene, inserting it into a suitable vector, transforming host cells, selecting transformed cells, culturing transformed clones, and extracting and purifying the recombinant product.

    Cloning Vectors

    • A cloning vector is a DNA molecule that carries a foreign DNA into a host cell (usually bacterial or yeast).
    • Cloning vectors have the ability to replicate, producing many copies of themselves along with the foreign DNA.
    • Types of cloning vectors include plasmids, phages, cosmids, bacterial artificial chromosomes (BAC), and yeast artificial chromosomes (YAC).

    Plasmids

    • Plasmids are small, circular, extrachromosomal DNA molecules found in bacteria that can replicate on their own outside of a host cell.
    • Plasmids have a cloning limit of 100 to 10,000 base pairs or 0.1-10 kilobases (kb).
    • Plasmids can confer advantages to host cells, such as antibiotic resistance, antibiotic production, sugar fermentation, degradation of aromatic compounds, heavy metal resistance, and toxin production.

    Phages

    • Phages are derivatives of bacteriophage lambda (λ phage), a virus that infects E. coli.
    • Phages have a linear DNA molecule that can be replaced with foreign DNA without disrupting its life cycle.
    • Phages have a high transformation efficiency, about 1000 times greater than that of plasmid vectors.
    • Phages have a larger cloning limit than plasmids, consisting of 8-25 kb.

    Cosmids

    • Cosmids are extrachromosomal circular DNA molecules that combine features of plasmids and phages.
    • Cosmids have a high transformation efficiency and a larger cloning limit than plasmids or phages, consisting of 35-50 kb.

    Bacterial Artificial Chromosomes (BAC)

    • BACs are based on bacterial plasmids and have a cloning limit of 75-300 kb.
    • Examples of BACs include F-plasmid and P1-plasmid.

    Yeast Artificial Chromosomes (YAC)

    • YACs are artificial chromosomes that replicate in yeast cells.
    • YACs are hybrids of bacterial plasmid DNA and yeast DNA.
    • YACs have a cloning limit of 100-1000 kb, but have very low efficiency.

    Microorganisms as Expression Hosts

    • Microorganisms are used as expression hosts for the production of recombinant products, such as proteins, enzymes, and products of multi-enzyme pathways.
    • Host requirements include suitable vectors, effective transformation, and ready fermentation and scale-up capacity.

    Microbial Expression Hosts

    • Examples of microbial expression hosts include Escherichia coli (E. coli), Saccharomyces cerevisiae, and Bacillus spp.
    • E. coli has advantages such as numerous specific vectors, high transformation efficiencies, and well-understood fermentations, but has disadvantages such as low expression yields.
    • Saccharomyces cerevisiae has advantages such as moderate expression yields and well-understood fermentations, but has disadvantages such as low-medium transformation efficiency and limited range of vector systems.
    • Bacillus spp. has advantages such as high transformation efficiencies, high expression yields, and simple fermentations, but has disadvantages such as limited range of vector systems.

    Expression Development

    • Expression development involves the use of multicopy plasmids, multiple selective markers, different promoters, multiple promoters, multiple ori sequences, and N-terminal tags.

    Viral Vectors

    • Viral vectors are viruses that carry a modified or foreign gene and are commonly used in gene therapy.
    • Examples of viruses used as vectors in gene therapy include retroviruses, adenoviruses, parvoviruses, herpesviruses, and poxviruses.

    Contribution of Microbiology to Biotechnology

    • Microbiology contributes to biotechnology by supplying antibiotics, cloning process, industrial enzymes, industrial products, food and beverages industries, research enzymes, medicine, and agriculture.

    Agrobacterium in Biotechnology

    • Agrobacterium is a bacteria that causes tumours in plants and has the ability to transfer genes to plants.
    • The tumour-inducing (Ti) plasmid is responsible for the production of the tumour and the transforming abilities.
    • The plasmid is 'disarmed' by deletion of the tumour-inducing genes, and the T-DNA is transferred and inserted into the host chromosome.

    Golden Rice

    • Golden rice is a genetically modified rice that produces beta-carotene, a precursor to vitamin A, to combat vitamin A deficiency.

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    LU3 Microbiology sem 2 2022.pdf

    Description

    This quiz covers the basic principles of modern biotechnology, including gene cloning, biochemistry, genetics, microbiology, and cell biology. It focuses on the general strategy of biotechnology using microbiology.

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