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Emily Summers Engineering Case Studies Human Insulin 2.2.14
If you cannot produce the hormone insulin, you suffer from type 1 diabetes mellitus.
Insulin used to be extracted from pancreatic tissue of pigs that had been
slaughtered, however this was ineffective and expensive because only a small
amount of insulin is produced
Scientists used centrifugation to separate mRNA of the correct length from
On finding this they used reverse transcriptase enzymes to synthesise a
complementary DNA strand.
The addition of DNA polymerase and DNA nucleotides means the second strand is
built on using this copied DNA as a template, producing a cDNA gene
unpaired nucleotides are added at ends to give complementary sticky ends to
those on the cut plasmid
Plasmids are cut open with restriction enzymes and mixed with cDNA, some
plasmids take up the gene
DNA lligase seals up the plasmids which are recombinant
These plasmids are mixed with bacteria, and some take up the recombinant plasmids
The bacteria are grown on an agar plate where the bacteria cells grow to produce a
mound of cloned cells, a colony.