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Biotechnology: Principles and Processes — BIOLOGY STD 12 Science — Question

Gujarat BoardEnglish MediumSTD 12 ScienceBIOLOGYBiotechnology: Principles and Processes5 Marks
Question
Explain the different steps in the construction of recombinant DNA.
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Answer

Isolation of the genetic material (DNA):
  • RNA is removed by treatment with ribonuclease and proteins are removed by treatment with protease.
  • After several treatments, the purified DNA is precipitated by adding chilled ethanol.
  • The bacterial/ plant/ animal cell is broken down by enzymes to release DNA, along with RNA, proteins, polysaccharides and lipids.
  • Bacterial cell is treated with enzyme lysozyme.
  • Plant cell is treated with enzyme cellulase.
  • Fungal cell is treated with chitinase.
Cutting of DNA at specific locations:
  • The DNA is cut using restriction enzymes.
  • The purified DNA is incubated, with the specific restriction enzyme at conditions optimum for the enzyme to act. Isolation of desired DNA fragment
  • Using agarose gel electrophoresis, the activity of the restriction enzymes can be checked.
  • Since the DNA is negatively charged, it moves towards the positive electrode or anode and in the process, DNA fragments separate out based on their sizes.
  • The desired DNA fragment is eluted out.
Amplification of gene of interest using PCR:
  • The Polymerase Chain Reaction (PCR) is a reaction in which amplification of specific DNA sequences is carried out in vitro.

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