PROCESSES OF RECOMBINANT DNA TECHNOLOGY
- Recombinant DNA ( rDNA ) technology involves the following stages
- Nucleic acid (DNA or RNA) is the genetic material of all organisms.
- It is DNA in majority organisms.
- For cutting the DNA with restriction enzymes it needs to be pure and free from other macromolecules.
- Because the DNA is covered by the membranes,it has to break the cell open to release DNA and other macromolecules like RNA, proteins, polysaccharides and lipids.
- It is obtained by treating the bacterial cells/plant or animal tissue with enzymes such as lysozyme (bacteria), cellulase (plant cells), chitinase (fungus).
- As we know that genes are present on long molecules of DNA interwined with proteins like histones, the RNA can be removed by treating with ribonuclease while proteins can be removed by treating with protease Other molecules are removed by proper treatments.
- The purified DNA finally precipitates out after the addition of chilled ethanol.
- This is seen as collection of fine threads in the suspension.
- Under the optimal conditions, the purified DNA is cut by the restriction enzyme.
- Agarose gel electrophoresis is used to check the progress of a restriction enzyme digestion.
- Since DNA is a negatively charged molecule, it moves towards the positive electrode (anode).
- This process is also repeated with the vector
- Formation of Recombinant DNA (rDNA)
- After the cutting of the source DNA and the ector DNA with a specific restriction enzyme, the cut out gene of interest from the source DNA and the cut vector with space are mixed and ligase enzyme is added.
- This results in the formation of a rDNA or hybrid DNA chimeric DNA .
- Continue to Read......... In Part - 2 article
========================================
Please do not enter any spam link or word in the comment box