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Question
Cloning of genes, play a very significant role in genetic engineering, helping the transfer of desirable foreign genes into different hosts. The scientists, to make this process easier and effective are creating engineered vectors in such a way that they help easy linking of foreign DNA and selection of recombinants from non-recombinants. 'pBR322' is one such engineered vectors developed by scientists. A diagram of an engineered vector pBR322 is given below:
- Name the host for this cloning vector.
- Identify 'Rop' and 'Ori' in the diagram from 'U', 'V', 'W', 'X', 'Y' and 'Z'. Write their functions.
- Draw the fragments that will be formed by the action of 'Z' (marked in the diagram) on the specific site of the DNA segment given below:
5' --- GTACGAATCCTGA --- 3'
3' --- CATGCTTAGGACT --- 3'
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Solution
- The host for this cloning vector is E.coli.
- Rop is W and Ori is U.
Function of Rop = to provide coding for plasmid replication-related proteins.
Function of Ori = Any DNA fragment can cause the host cells to reproduce any other DNA fragment when linked to this sequence, which is where replication begins. - Z is EcoRI. The recognition sequence's 5' terminus contains a cut between G and A bases. -GAATTC-3' in 5'. Therefore, the fragments produced if the given sequence is treated with EcoRI are:
5' --- GTACG AATTCCTGA --- 3'
3' --- CATGCTTAA GGACT --- 5'
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RELATED QUESTIONS
Explain the mechanism of action of restriction endonucleases that makes them suitable for genetic engineering.
ECoRI is obtained from ______.
Match the Column I (Restriction enzyme) with Column II (Recognition sequence) and select the correct option.
| column I (Restriction Enzyme) | column II (Recognition sequence) | ||
| i. | Alu | P. | 5'---G- ↓-G-A-T-C-C---3' 3'---C-C-T-A-G- -G---5 |
| ii. | BamHI | Q. | 5'--- A-G- ↓ -C-T---3' 3' ---T-C- G-A--- 5' |
| iii. | Eco RI | R. | 5'--G-T-C- ↓ -G-A-C--3' I 3'--C-A-G-t-C-T-G--5' |
| iv. | HindII | S. | 5'---G- ↓ -A-A-T-T-C---3' 3'---C-T-T-A-A- -G---5' |
Which of the following DNA sequence is recognised and cut by EcoR I?
Identify the process by which bacteria protect their own DNA from the attack of restriction enzymes.
Which molecules are called molecular scissors? Why are they called so?
Which of the following statements is correct with respect to cellular respiration?
Nucleic acid segment tagged with a radioactive molecule is called ______.
A recombinant DNA molecule was created by ligating a gene to a plasmid vector. By mistake, an exonuclease was added to the tube containing the recombinant DNA. How does this affect the next step in the experiment i.e. bacterial transformation?
Importance of PCR.
