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Question
What is gene cloning? Explain different tools used for it.
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Solution
- Gene cloning is a process which involves the isolation, transfer of a specific sequence of DNA through a vector and amplification in a host organism.
- There are three types of biological tools used in rDNA technology viz. enzymes, cloning vectors (vehicle DNA) and competent host (cloning organisms) for transformation with recombinant DNA.
RELATED QUESTIONS
Give the applications of the PCR technique.
The bacterium which causes a plant disease called crown gall is _______________.
A PCR machine can raise the temperature up to 100°C but after that, it is not able to lower the temperature below 70°C automatically. Which step of PCR will be hampered first in this faulty machine? Explain why?
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' |
The most important feature in a plasmid to serve as a vector in gene cloning experiment is ______.
Name a recombinant vaccine that is currently being used in vaccination programme.
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?
Restriction enzymes were discovered by ______.
The structure below shows pUC18, which is similar to pBR322 in its function. However, they differ in some of their restriction sites and the number of ori. The ori number for pBR322 is approximately 20.
- How are puc18 and pBR322 used in biotechnological studies?
OR
What will be the impact if the ori in the above structure gets damaged? - The lac z gene has many recognition sites. Study the segment of DNA given below and answer the questions.
5’... ATC GTA AAG CTT CAT…3’
3’... TAG CAT TTC GAA GTA…5’
i) Applying your knowledge of palindrome sequences identify and mark the possible region where the restriction enzyme X will act.
ii) Restriction enzyme Y was used to extract a gene of interest from a plant. This gene needs to be inserted in the given DNA segment which has been treated with restriction enzyme X. Will there be a successful recombination? Explain with a reason. - Which one of the two (pUC18 and pBR322) would you prefer for biotechnological studies? Justify.
Tissue plasminogen activator and urokinase are used in the treatment of ______.
