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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.
Solution :
1. Theoretical Foundation of Restriction Endonucleases
Restriction endonucleases, often termed "molecular scissors," are enzymes capable of recognizing specific base pair sequences known as palindromic sequences within a DNA molecule. Upon recognition, they hydrolyze the phosphodiester backbone of both strands of the DNA duplex. The specific site of action is the Recognition Site [Per the mechanism of Type II Restriction Endonucleases].
2. Comparative Analysis of DNA Cleavage Components
| Component | Description |
|---|---|
| Restriction Enzyme | $EcoRI$ (Isolated from Escherichia coli RY13). |
| Substrate DNA | Double-stranded DNA containing the palindrome $5'-GAATTC-3'$. |
| Recognition Site | The hexanucleotide sequence: $5'-GAATTC-3' / 3'-CTTAAG-5'$. |
| Product | DNA fragments with single-stranded overhanging ends known as "Sticky Ends." |
3. Diagrammatic Representation of $EcoRI$ Action
4. Sequential Mechanism of Action
- Step 1: Scanning – The $EcoRI$ enzyme binds to the DNA molecule and scans the length of the phosphodiester backbone until it encounters the specific recognition sequence [By the principle of facilitated diffusion and hydrogen bond probing].
- Step 2: Cleavage – The enzyme catalyzes the hydrolysis of the phosphodiester bond between the Guanine ($G$) and Adenine ($A$) bases on both strands [Per the chemical mechanism of nucleophilic attack facilitated by $Mg^{2+}$ ions].
- Step 3: Dissociation – The enzyme releases the two DNA segments. Because the cut is staggered, each fragment possesses an overhanging single-stranded tail ($5'-AATT-3'$). These are referred to as sticky ends, which facilitate hydrogen bonding with complementary sequences on other DNA fragments [Base-pairing logic via Chargaff’s Rules].
Final Solution: The action of the restriction enzyme EcoRI on the substrate DNA results in the cleavage of the $5'-GAATTC-3'$ sequence between the $G$ and $A$ residues, producing two distinct DNA fragments characterized by protruding, complementary single-stranded sticky ends.
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.
- 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(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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