Genetic Engineering

Making Recombinant DNA

• Specification Point 5.12
• Understand how restriction enzymes are used to cut DNA at specific
sites and ligase enzymes are used to join pieces of DNA together
• Genetic engineering is changing the traits of one organism by
inserting genetic material from another organism
• The organism receiving the genetic material is said to be ‘genetically
modified’
• The DNA of the organism that now contains DNA from another
organism as well is known as ‘recombinant DNA’
• The process is as follows:
• The gene that is to be inserted is located in the original organism (for
example, this could be the gene for human insulin)
• Restriction enzymes are used to isolate the required gene, leaving it
with ‘sticky ends’ (a short section of unpaired bases)
• A vector, which is usually a bacterial plasmid or a virus, is cut by the
same restriction enzyme leaving it with corresponding sticky ends
• The vector and the isolated gene
are joined together by ligase
enzyme
• If two pieces of DNA have
matching sticky ends (because
they have been cut by the same
restriction enzyme), ligase will
link them to form a single,
unbroken molecule of DNA
Vectors for Recombinant DNA
• Specification Point 5.13
• Understand how plasmids and viruses can act as vectors, which take up
pieces of DNA, and then insert this recombinant DNA into other cells

•A vector is a vehicle to artificially carry foreign genetic material into another cell, where it can be
replicated and/or expressed
•The two main types of vectors are bacterial plasmids and viruses
• Plasmids are circles of DNA that are found
in bacteria
• When the bacteria reproduce the plasmids
are copied as well and so a recombinant
plasmid can quickly be spread as the
bacteria multiply and they will then all
express the gene
• Viruses will carry recombinant DNA
and infect organisms that they come into
contact with, making the host organism
express the recombinant DNA
Manufacture of Insulin
Specification Point 5.14
Understand how large amounts of human insulin can be manufactured from genetically modified bacteria that are grown in a fermenter

• Insulin is needed in large

quantities to help treat diabetes
• Prior to the development of
genetic engineering, insulin for
human use was collected from
pigs
• Bacteria are now genetically
engineered to produce human
insulin which can be collected
and purified for use with no fear
of rejection
• Once some bacteria have been genetically engineered they are grown
in large quantities in a fermenter
• Each bacterial cell will produce a tiny mass of insulin
• In this way large quantities of insulin protein can be produced,
extracted and purified for use by diabetics
Improving food production
• Genetically modified plants are plants that have had foreign DNA
inserted into their genome
• This is usually done to improve food production in some way
• For example:
• Tomatoes have been genetically modified to make them grow larger fruit
• Wild rice has been genetically modified to produce beta carotene (needed by
humans to make vitamin A)
• Crop plants have been genetically modified to be resistant to herbicides
Transgenic Organisms
• The term ‘transgenic’ means the transfer of genetic material from
one species to a different species
• This means that a transgenic organism contains genes from a different
species
• For example, the recombinant bacteria used to  produce human insulin
are transgenic as they contain genetic material from a human