Question

Describe the basic steps of Recombinant DNA Technology.

Hint:

Recombinant DNA Technology combines genes and vectors using restriction enzymes and ligase, followed by transformation and selection in host cells.

Answer 1

Recombinant DNA Technology involves combining DNA molecules from different sources to create new genetic combinations. The basic steps are:

1. Isolation of DNA: Extract the gene of interest from the donor organism and the vector DNA (usually a plasmid).
2. Cutting DNA: Use restriction enzymes (restriction endonucleases) to cut both the gene and the vector DNA at specific sequences, creating compatible ends.
3. Ligation: Join the gene and vector DNA fragments using the enzyme DNA ligase to form a recombinant DNA molecule.
4. Transformation: Introduce the recombinant DNA into a suitable host cell (like E. coli) by transformation.
5. Selection: Grow the host cells on selective media to identify cells that have taken up the recombinant DNA.
6. Screening: Confirm the presence and expression of the inserted gene in transformed cells.

This technology allows production of genetically modified organisms, therapeutic proteins, and vaccines.