Question
Enlist and write in brief about the different biological tools required in r-DNA technology.
✓
Answer
The biological tools used in r-DNA technology are various enzymes, cloning vectors and competent hosts.
(1) Enzymes:
(1) Enzymes:
- Enzymes like lysozymes, nucleases (exonucleases and endonucleases), DNA ligase, reverse transcriptase, DNA polymerase, alkaline phosphatases, etc. are used in r-DNA technology.
- The restriction endonucleases are used as biological or molecular scissors. They are able to cut a DNA molecule at a specific recognition site.
(2) Vectors:
- Vectors are DNA molecules which carry foreign DNA segment and replicate inside the host cell.
- Vectors may be plasmids, bacteriophages (M13, lambda virus), cosmid, phagemids, BAC (bacterial artificial chromosome), YAC (yeast artificial chromosome), transposons, baculoviruses and mammalian artificial chromosomes (MACs).
- Most commonly used vectors are plasmid vectors (pBR 322, pUC, Ti plasmid) and bacteriophages (lamda phage, M13 phage).
(3) Competent host cells:
- They are bacteria like Bacillus haemophilus, Helicobacter pyroliand E. coli.
- Mostly E. coli is used for the transformation with recombinant DNA.
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