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प्रश्न
Enlist different types of restriction enzymes commonly used in rDNA technology? Write about their role.
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उत्तर
There are three types of restriction enzymes:
- Type I – These enzymes function simultaneously as endonuclease and methylase e.g. EcoK.
- Type II – These enzymes have separate activities for cleaving and methylation; they are more stable and are used in rDNA technology e.g. EcoRI, BglII; these enzymes cut DNA at specific sites within the palindrome. There are thousands of type II restriction enzymes that are recognized/ discovered.
- Type III – These enzymes cut DNA at specific non-palindromic sequences e.g. HpaI, MboII.
Role of restriction enzymes:
Restriction enzymes either cut straight across the DNA in the region of palindrome to give blunt ends or cuts producing short, single-stranded projections at each end of DNA to produce, cohesive or sticky ends or staggered ends.
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संबंधित प्रश्न
Rearrange the following in the correct sequence to accomplish an important biological reaction:
(a) In vitro synthesis of copies of DNA of interest
(b) Chemically synthesized oligonucleotides
(c) Enzyme DNA-polymerase
(d) Complementary region of DNA
(e) Genomic DNA template
(f) Nucleotides provided
(g) Primers
(h) Thermostable DNA-polymerase (from Thermus aquaticus)
(i) Denaturation of ds-DNA
The enzyme nuclease hydrolyses ______ of polynucleotide chain of DNA.
What is the role of Taq-polymerase in PCR technology?
Which of the following DNA sequence is recognised and cut by EcoR I?
Identify the CORRECT order of steps in Polymerase Chain Reaction (PCR).
What is meant by gene cloning?
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?
Identify the desirable characteristics for a plasmid used in rDNA technology from the following.
- Ability to multiply and express outside the host in a bioreactor.
- A highly active promoter.
- A site at which replication can be initiated.
- One or more identifiable marker genes.
- One or more unique restriction sites.
The polymerase chain reaction is a powerful technique to ______.
In vitro amplification of DNA segments is called______.
