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Questions and Answers
What is the primary function of Agrobacterium in biotechnology?
What is the primary function of Agrobacterium in biotechnology?
Which of the following is NOT a contribution of microbiology to biotechnology?
Which of the following is NOT a contribution of microbiology to biotechnology?
What is the name of the disease caused by Agrobacterium in plants?
What is the name of the disease caused by Agrobacterium in plants?
What is the role of T-DNA in Agrobacterium?
What is the role of T-DNA in Agrobacterium?
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What is the purpose of 'disarming' the Ti plasmid?
What is the purpose of 'disarming' the Ti plasmid?
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What is the size of the circular DNA plasmid responsible for tumour production and gene transfer?
What is the size of the circular DNA plasmid responsible for tumour production and gene transfer?
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Which of the following viruses is used as a vector in gene therapy?
Which of the following viruses is used as a vector in gene therapy?
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What is the significance of Golden rice?
What is the significance of Golden rice?
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What is the role of border repeats in T-DNA?
What is the role of border repeats in T-DNA?
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Which of the following is an application of microbiology in agriculture?
Which of the following is an application of microbiology in agriculture?
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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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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.
