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’
Restriction enzymes cut DNA strands
 The DNA of the organism that now
unequally to form ‘sticky ends’
contains DNA from another organism
as well is known as ‘recombinant
DNA’  The vector and the isolated gene are
joined together by ligase enzyme
The process is as follows:  If two pieces of DNA have matching
sticky ends (because they have been
cut by the same restriction enzyme),
 The gene that is to be inserted is
ligase will link them to form a single,
located in the original organism (for
unbroken molecule of DNA
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
DNA ligase is used to join two separate pieces from genetically modified bacteria that
of DNA together are grown in a fermenter
 Insulin is needed in large quantities to
help treat diabetes
Vectors for Recombinant DNA
 Prior to the development of genetic
Specification Point 5.13
engineering, insulin for human use was
 Understand how plasmids and viruses collected from pigs
can act as vectors, which take up pieces
 Bacteria are now genetically
of DNA, and then insert this
engineered to produce human
recombinant DNA into other cells
insulin which can be collected and
 A vector is a vehicle to artificially purified for use with no fear of rejection
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

Vectors can be bacterial plasmids or viruses

 Plasmids are circles of DNA that are
found in bacteria
The process of creating human insulin by
 When the bacteria reproduce genetic engineering
the plasmids are copied as well and so
 Once some bacteria have been
a recombinant plasmid can quickly be
genetically engineered they are grown in
spread as the bacteria multiply and they
will then all express the gene large quantities in a fermenter
 Each bacterial cell will produce a tiny
 Viruses will carry recombinant DNA
and infect organisms that they come mass of insulin
into contact with, making the host  In this way large quantities of insulin
organism express the recombinant protein can be produced, extracted and
DNA purified for use by diabetics
Manufacture of Insulin
Specification Point 5.14
 Understand how large amounts of
human insulin can be manufactured