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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
Identify desired gene
Restriction enzyme cuts DNA & isolates gene
Same restriction enzyme cuts vector
Splice gene with vector
DNA ligase bind sticky ends
Form recombinant DNA
Insert into host cell
Cultured + cloned
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