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प्रश्न
Explain what are the desirable characteristics of an ideal cloning vector used in rDNA technology.
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उत्तर
Desirable characteristics of an ideal cloning vector:
- Origin of replication: It is a specific sequence of nucleotides that serves as a binding site for initiator protein (Helicase) to start replication. Any foreign DNA linked with vector DNA also gets replicated along with it, hence it is important for maintaining the copy number of foreign DNA.
- Selectable markers: Cloning vectors must contain at least one selectable marker or it helps in:
– detection of transformed host cells,
– Identification and elimination of nontransformed post cells,
– Isolation and growth of transformed host cells. - Cloning sites: A cloning site is a restriction endonuclease recognition site or target site in vector DNA. It has a palindromic sequence of nitrogenous bases cut by a specific restriction endonuclease enzyme for insertion and linking of foreign DNA.
- Small size: A vector should be small in size because large molecules bind is break up during isolation and purification.
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संबंधित प्रश्न
What would be the molar concentration of human DNA in a human cell?
Give one significant difference between:
Electroporation and Gene Gun.
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i. Capability of science to change the genetic material for getting new specific products through rDNA technology, polymerase chain reaction (PCR), microarrays, cell culture and fusion, and bioprocessing.
ii. Ownership of technology and its sociopolitical impact.
iii. Modem biotechnology makes use of only fermentation technology.
iv. Classical biotechnology relies only on principles of plant and animal tissue culture.
____________ cuts the DNA within the specific positions.
What is meant by gene cloning?
Observe the given sequence of nitrogenous bases on a DNA fragments and answer the following questions:
5' - CAGAATTCTTA - 3'
3' - GTCTTAAGAAT - 5'
- Name a restriction enzyme which can recognise this DNA sequence.
- Write the sequence after digestion.
- Why are the ends generated after digestion called sticky ends?
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.
Study the diagram given below and answer the questions that follow.
- Name the cloning vector shown above. In which organism in this cloning vector inserted?
-
Mention any two restriction sites shown in the diagram.
- Name any two selectable markers shown in the diagram.
