Recombinant DNA Technology
• Class 12 Biology / Biotechnology
• Detailed PPT Presentation
Introduction to rDNA
• Recombinant DNA (rDNA) technology is a
technique to join DNA from two different
sources.
• The resulting DNA is inserted into a host
organism to express the desired gene.
• Used widely in medicine, agriculture,
research, and industry.
Tools of rDNA Technology
• - Restriction Enzymes: Cut DNA at specific
sites.
• - DNA Ligase: Joins DNA fragments.
• - Vectors: Carry foreign DNA (e.g., plasmids,
viruses).
• - Host Cells: Organisms into which
recombinant DNA is inserted.
• - Selectable Markers: Help identify
transformed cells.
Steps in rDNA Technology –
Overview
• 1. Isolation of genetic material (DNA)
• 2. Cutting DNA at specific sites using
restriction enzymes
• 3. Amplification of gene of interest using PCR
• 4. Insertion of DNA into vector
• 5. Introduction into host
• 6. Selection of recombinants
• 7. Expression and purification of product
Isolation of DNA & Restriction
Digestion
• DNA is extracted from donor and vector cells.
• Restriction enzymes cut DNA at specific
sequences to obtain target gene and prepare
vector.
PCR – Amplification of Gene of
Interest
• Polymerase Chain Reaction (PCR) makes
millions of copies of the gene.
• Steps:
• 1. Denaturation
• 2. Annealing
• 3. Extension
• Components: DNA template, primers, DNA
polymerase, dNTPs.
Ligation – Joining DNA with Vector
• Cut vector and gene are joined using DNA
ligase.
• Forms recombinant DNA molecule.
Introduction into Host Cell
• The recombinant plasmid is introduced into a
host cell (e.g., E. coli).
• Techniques: Heat Shock, Electroporation,
Microinjection, Gene Gun.
Selection and Screening of
Recombinants
• Not all host cells take up recombinant DNA.
• Selectable markers like antibiotic resistance
are used.
• Only transformed cells grow on selective
media.
Expression and Purification of
Gene Product
• Host cell expresses the inserted gene to
produce protein.
• The protein is then extracted and purified.
• Used for medicine, research, etc.
Applications of rDNA Technology
• - Medicine: Insulin, vaccines, gene therapy
• - Agriculture: GM crops (Bt cotton, Golden
Rice)
• - Industry: Enzymes (amylase, cellulase)
• - Environment: Bioremediation, oil spill
treatment
Conclusion
• Recombinant DNA technology has
revolutionized biotechnology.
• It enables targeted genetic manipulation for
beneficial outcomes.
• Its applications continue to grow in every field
of life sciences.

Recombinant_DNA_Technology_Class12.pptxok

  • 1.
    Recombinant DNA Technology •Class 12 Biology / Biotechnology • Detailed PPT Presentation
  • 2.
    Introduction to rDNA •Recombinant DNA (rDNA) technology is a technique to join DNA from two different sources. • The resulting DNA is inserted into a host organism to express the desired gene. • Used widely in medicine, agriculture, research, and industry.
  • 3.
    Tools of rDNATechnology • - Restriction Enzymes: Cut DNA at specific sites. • - DNA Ligase: Joins DNA fragments. • - Vectors: Carry foreign DNA (e.g., plasmids, viruses). • - Host Cells: Organisms into which recombinant DNA is inserted. • - Selectable Markers: Help identify transformed cells.
  • 4.
    Steps in rDNATechnology – Overview • 1. Isolation of genetic material (DNA) • 2. Cutting DNA at specific sites using restriction enzymes • 3. Amplification of gene of interest using PCR • 4. Insertion of DNA into vector • 5. Introduction into host • 6. Selection of recombinants • 7. Expression and purification of product
  • 5.
    Isolation of DNA& Restriction Digestion • DNA is extracted from donor and vector cells. • Restriction enzymes cut DNA at specific sequences to obtain target gene and prepare vector.
  • 6.
    PCR – Amplificationof Gene of Interest • Polymerase Chain Reaction (PCR) makes millions of copies of the gene. • Steps: • 1. Denaturation • 2. Annealing • 3. Extension • Components: DNA template, primers, DNA polymerase, dNTPs.
  • 7.
    Ligation – JoiningDNA with Vector • Cut vector and gene are joined using DNA ligase. • Forms recombinant DNA molecule.
  • 8.
    Introduction into HostCell • The recombinant plasmid is introduced into a host cell (e.g., E. coli). • Techniques: Heat Shock, Electroporation, Microinjection, Gene Gun.
  • 9.
    Selection and Screeningof Recombinants • Not all host cells take up recombinant DNA. • Selectable markers like antibiotic resistance are used. • Only transformed cells grow on selective media.
  • 10.
    Expression and Purificationof Gene Product • Host cell expresses the inserted gene to produce protein. • The protein is then extracted and purified. • Used for medicine, research, etc.
  • 11.
    Applications of rDNATechnology • - Medicine: Insulin, vaccines, gene therapy • - Agriculture: GM crops (Bt cotton, Golden Rice) • - Industry: Enzymes (amylase, cellulase) • - Environment: Bioremediation, oil spill treatment
  • 12.
    Conclusion • Recombinant DNAtechnology has revolutionized biotechnology. • It enables targeted genetic manipulation for beneficial outcomes. • Its applications continue to grow in every field of life sciences.