BIOTECHNOLOGY- PROCESSES OF RECOMBINANT DNA TECHNOLOGY

 THE STEPS INVOLVED IN RECOMBINANT DNA TECHNOLOGY ARE;

1. ISOLATION OF DNA / GENETIC MATERIAL:

Bacterial cells can be lysed using lysozyme, plant cells can be treated with cellulase and fungal cells can be treated with chitinase to break open cells and release DNA and cell content. 

RNA can be removed by treating with RNase and proteins are removed by treating with proteases. Purified DNA can be precipitated with chilled ethanol. Cloudy, precipitated DNA can be observed in test tube.

2. CUTTING DNA USING RESTRICTION ENDONUCLEASES:

DNA from the source organism is cleaved by restriction endonuclease to create restriction fragments.

Vector DNA is also cleaved using the same restriction endonuclease.

These fragments can be separated using agarose gel electrophoresis. 

3. INSERTION OF DESIRED FOREIGN GENE INTO CLONING VECTOR:

Gene of interest is joined with vector DNA using DNA ligase. The combination of vector DNA and foreign DNA is called RECOMBINANT DNA OR CHIMERIC DNA. 

4. TRANSFER OF rDNA INTO HOST:

This can be achieved by a transformation in bacterial cells or Microinjection in animal cells or by using gene gun [ biolistics ] in plant cells.