Genetic Engineering

Plasmid
circular piece of DNA
Gene for antibiotic Gene for the enzyme In practice the process is not always successful and three
resistence beta galactosidase possible types of bacteria can be produced. It is
important only to grow bacteria which have successfully
been transformed and they have been transformed with a
plasmid has taken up the new gene. Culturing any other
bacteria would reduce the potential product yield.

Restriction
endonuclease
binding site
A plasmid is created with two marker genes. 5
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The plasmid is mixed with a restriction endonuclease which This bacteria has not been transformed and lacks the
cuts the DNA across the beta galactosidase gene. This creates engineered plasmid. As a consequence it lacks the antibiotic
sticky ends, short single stranded lengths of DNA either side resistence gene and will not grow in a medium containing
of the break which are complementary to each other. the antibiotic. This leaves you with the other two types.

New Gene

Sticky ends
I

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The gene to be cloned is added to the plasmid and base pairing A bacteria that is transformed but with a plasmid that did not
occurs between the complementary sticky ends. DNA ligase is take up the gene.. As a consequence the galactosidase gene is
also added which covalently bonds the foreign gene into the not inactivated and will continue to produce an enzyme
plasmid a process known as annealing. enabling the bacteria to grow in a medium containing lactose.

II
New Gene

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The next step is to place the recombinant DNA plasmid into a These bacteria have been succesfully transformed containing
suitable host cell such as a bacteria this may be done in a variety the marker gene for antibiotic resistence and aslo have the
of ways for example by using viral vectors. This process is new gene. The new gene inactivates the galactosidase gene.
called transformation.

III

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