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what is recombinant DNA technology?
joining together pieces of DNA from different organisms and inserting into host organism to produce proteins
what is a plasmid
small circular extrachromosomal piece of double stranded DNA naturally found in many bacteria
four key elements of plasmids
origin of replication
antibiotic resistance gene
promoter
selectable marker
origin of replication
allow initiation of replication using DNA polymerase
antibiotic resistance gene
allows selection of cells containing the plasmid (antibiotic is included in control media)
promoter
drives transcription of your gene in cells with appropriate transcription factors
selectable marker
colour-based selection of cells that have taken up the recombinant vector
how to cut and paste DNA into plasmids
using restriction enzymes that recognise specific sequences - uses sticky ends so new gene can be attached
universal genetic code
all living organisms use the same genetic code (same bases code for same amino acids)
universal genetic code significance
bacterial cells are able to correctly transcribe and translate human genes
what can prokaryotes not do in gene expression
cannot splice intons from pre-mRNA so resulting protein would be nonsense
cDNA
before gene is inserted into plasmid, introns are removed through reverse transcription
also makes size more manageable
