recombinant dna technology Flashcards
(20 cards)
What does recombinant DNA technology involve?
Transfer of fragments of DNA from one organism, or species to another.
Why can this be done between different species?
Genetic code is universal.
What is a transgenic organism?
Organism that’s received transferred DNA.
How can the fragments be obtained?
Reverse transcriptase, restriction endonucleases, a gene machine.
How is reverse transcriptase used?
Uses mRNA which is removed from a cell.
What is the first step in using reverse transcriptase?
mRNA is used as a template to produce required gene from DNA.
What is the next step after using mRNA as a template?
mRNA is mixed with free DNA nucleotides and the enzyme reverse transcriptase.
What is the next step after mixing mRNA with DNA nucleotides?
Free DNA nucleotides align next to their complimentary bases.
What is the next step after aligning DNA nucleotides?
Reverse transcriptase joins the DNA nucleotides together to produce a fragment of DNA.
What is the DNA produced from this technique?
Complementary DNA (cDNA).
What is produced from cDNA?
Double stranded DNA using DNA nucleotides and DNA polymerase.
What does it mean if there are no introns in the fragments?
The fragments can be transcribed by bacteria.
What will fragments using restriction enzymes contain?
Introns (if from a eukaryotic organism).
What does the gene machine enable?
Production of fragments without needing pre-existing DNA or mRNA as a template.
What can be used as a template in the gene machine?
Amino acid sequence of a protein.
What does the gene machine produced fragments not contain?
Introns.
What are promoter and terminator regions?
Sections of DNA which must be added to the fragment for successful transcription.
How can fragments be amplified?
In vivo, in vitro.
What does in vivo mean?
Where the copies are made inside a living organism.
What does in vitro mean?
Where the copies are made outside a living organism (usually by PCR).
