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Q10(a):
Describe briefly the following: (a) Origin of replication
Solution :
Definition and Functional Role
The Origin of Replication, commonly abbreviated as ori, is a specific DNA sequence located within a genome, plasmid, or viral chromosome where the process of DNA replication is initiated [Per the Semi-conservative model of DNA replication]. In the context of biotechnology and recombinant DNA technology, the ori serves as the critical regulatory switch that permits an extrachromosomal element (such as a plasmid) to replicate autonomously within a host cell.
Mechanism of Action
- Protein Recognition: Specific initiator proteins (e.g., DnaA in E. coli) recognize and bind to the conserved sequence of the ori site [Binding affinity is dictated by the chemical architecture of the DNA major groove].
- Strand Separation: Upon binding, the initiator proteins facilitate the local denaturation (unwinding) of the DNA double helix to create a replication bubble.
- Recruitment: This open configuration allows the assembly of the replication machinery, including DNA helicase, primase, and DNA polymerase, thereby commencing bidirectional or unidirectional synthesis [Molecular Biology of the Cell, Alberts et al.].
Role in Cloning Vectors
For a DNA fragment to be cloned successfully, it must be linked to an ori. The ori performs two primary roles in genetic engineering:
- Autonomous Replication: It ensures the foreign DNA is replicated independently of the host's chromosomal DNA.
- Copy Number Control: The ori sequence dictates the "copy number" of the plasmid—the number of copies of the plasmid maintained per host cell. Sequences that allow high copy numbers are highly desirable in biotechnology for maximizing the yield of recombinant protein or DNA product [Ref: Vector Biology Principles].
Visual Representation of Origin Function
Summary of Functional Characteristics
| Feature | Description |
|---|---|
| Sequence Composition | Often A-T rich regions to facilitate easier strand separation [Thermodynamic stability of A-T pairs is lower than G-C pairs]. |
| Initiation | Serves as the mandatory loading site for the pre-replication complex. |
| Biotech Utility | Essential for the survival and multiplication of recombinant vectors in host organisms. |
Final Solution: The Origin of Replication (ori) is a specific DNA sequence that acts as the initiation site for DNA synthesis, enabling the autonomous replication of vectors and determining the copy number of recombinant plasmids within a host system.
More Questions from Class 12 Biology Biotechnology: Principles and Processes EXERCISES
- Q1: Can you list 10 recombinant proteins which are used in medical practice? Find out where they are used as therapeutics (use the internet).
- Q1: Can you list 10 recombinant proteins which are used in medical practice? Find out where they are used as therapeutics (use the internet).
- Q2: Make a chart (with diagrammatic representation) showing a restriction enzyme, the substrate DNA on which it acts, the site at which it cuts DNA and the product it produces.
- Q2: Make a chart (with diagrammatic representation) showing a restriction enzyme, the substrate DNA on which it acts, the site at which it cuts DNA and the product it produces.
- Q3: From what you have learnt, can you tell whether enzymes are bigger or DNA is bigger in molecular size? How did you know?
- Q3: From what you have learnt, can you tell whether enzymes are bigger or DNA is bigger in molecular size? How did you know?
- Q4: What would be the molar concentration of human DNA in a human cell? Consult your teacher.
- Q4: What would be the molar concentration of human DNA in a human cell? Consult your teacher.
- Q5: Do eukaryotic cells have restriction endonucleases? Justify your answer.
- Q5: Do eukaryotic cells have restriction endonucleases? Justify your answer.
- Q6: Besides better aeration and mixing properties, what other advantages do stirred tank bioreactors have over shake flasks?
- Q6: Besides better aeration and mixing properties, what other advantages do stirred tank bioreactors have over shake flasks?
- Q7: Collect 5 examples of palindromic DNA sequences by consulting your teacher. Better try to create a palindromic sequence by following base-pair rules.
- Q7: Collect 5 examples of palindromic DNA sequences by consulting your teacher. Better try to create a palindromic sequence by following base-pair rules.
- Q8: Can you recall meiosis and indicate at what stage a recombinant DNA is made?
- Q8: Can you recall meiosis and indicate at what stage a recombinant DNA is made?
- Q9: Can you think and answer how a reporter enzyme can be used to monitor transformation of host cells by foreign DNA in addition to a selectable marker?
- Q9: Can you think and answer how a reporter enzyme can be used to monitor transformation of host cells by foreign DNA in addition to a selectable marker?
- Q10(a): Describe briefly the following: (a) Origin of replication
- Q10(b): Describe briefly the following: (b) Bioreactors
- Q10(b): Describe briefly the following: (b) Bioreactors
- Q10(c): Describe briefly the following: (c) Downstream processing
- Q10(c): Describe briefly the following: (c) Downstream processing
- Q11(a): Explain briefly (a) PCR
- Q11(a): Explain briefly (a) PCR
- Q11(b): Explain briefly (b) Restriction enzymes and DNA
- Q11(b): Explain briefly (b) Restriction enzymes and DNA
- Q11(c): Explain briefly (c) Chitinase
- Q11(c): Explain briefly (c) Chitinase
- Q12(a): Discuss with your teacher and find out how to distinguish between (a) Plasmid DNA and Chromosomal DNA
- Q12(a): Discuss with your teacher and find out how to distinguish between (a) Plasmid DNA and Chromosomal DNA
- Q12(b): Discuss with your teacher and find out how to distinguish between (b) RNA and DNA
- Q12(b): Discuss with your teacher and find out how to distinguish between (b) RNA and DNA
- Q12(c): Discuss with your teacher and find out how to distinguish between (c) Exonuclease and Endonuclease
- Q12(c): Discuss with your teacher and find out how to distinguish between (c) Exonuclease and Endonuclease
CBSE Solutions for Class 12 Biology Biotechnology: Principles and Processes
Chapters in CBSE - Class 12 Biology
- Reproduction in Organisms
- Human Reproduction
- Reproductive Health
- Principles of Inheritance and Variation
- Molecular Basis of Inheritance
- Evolution
- Human Health and Disease
- Strategies for Enhancement in Food Production
- Microbes in Human Welfare
- Biotechnology and Its Applications
- Organisms and Populations
- Ecosystem
- Biodiversity and Conservation
- Environmental Issues
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