Biotechnology

Recombinant DNA Technology

Process of combining DNA from different sources to produce artificial recombinant DNA

Recombinant DNA

Artificial DNA formed by combining DNA from different sources

Genetically Modified Organisms

Organisms with altered DNA

Transgenic Organisms

Organisms that have had their DNA altered with foreign DNA from different species

Process

1. Identify desired gene

2. Restriction enzyme cuts DNA & isolates gene

3. Same restriction enzyme cuts vector

4. Splice gene with vector

5. DNA ligase bind sticky ends

6. Form recombinant DNA

7. Insert into host cell

8. Cultured + cloned

9. Extracted & purified

Restriction Enzymes

Enzymes that cut DNA at specific sequences of nucleotides

Recognition Site

Specific sequence of nucleotides where enzyme cuts DNA

Palindromic

Sequences that reads same forwards & backwards

Straight Cut

Restriction enzyme makes clean break across 2 DNA strands to produce blunt ends

Blunt Ends

DNA fragments with paired base endings produced by straight cuts

Staggered Cut

Restriction enzyme cuts DNA at different locations to produce sticky ends

Sticky Ends

DNA fragments forming overhangs with unpaired base pairings produced by a staggered cut

DNA Ligase

Enzyme capable of combining 2 single stranded DNA segments into 1 structure

DNA Ligase Process

• Complementary bases MUST form hydrogen bonds

• Bind phosphate group to sugar molecule

Vectors

Bacterial plasmids/viral phages used to transfer DNA into a cell

Plasmid

Small circular strand of cytoplasmic DNA in bacteria that can replicate independently

Bacteriophages

Viruses that infect bacteria