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PRODUCING HUMAN INSULIN FROM BACTERIA

Why Bacteria?

Production and Collection of insulin

Advantages of using bacteria:

Insulin was originally extracted from pig and cow pancreases.

The bacteria are then grown on an industrial scale in huge fermenters to produce large amounts of insulin

• It is easier to create high quantities of insulin

• It can be produced more rapidly

• It is produced at lower a lower cost than animal insulin

• It is less likely to cause an adverse reaction as it’s identical to human insulin

• It overcomes ethical concerns from vegetarians and others

• This form of insulin is absorbed more rapidly

The insulin made is extracted from the fermenters and purified so that it can be sold and used.

This insulin is slightly different to human insulin (by a few amino acids) so it is far more effective to use bacteria to make actual human insulin

Growing the bacteria

Preparation of vector

Putting the vector into the bacteria

The bacteria with the recombinant plasmid is then grown to produce bacteria which can produce human insulin.

A DNA plasmid is removed from an E.coli bacterium

The plasmid is cut open using the same restriction enzymes as were used to cut out the gene of interest to give complementary sticky ends.

Isolation of human gene

The isolated gene then binds to the restriction site on the plasmid.

The "recipient bacteria" is either heat-shocked or it undergoes electroporation to make the membrane permiable enough to accept the plasmid

mRNA is taken from B cells from the human pancreas.

This mRNA is used to synthesise DNA which codes for insulin production

This creates a recombinant vector carrying the gene of interest.

mRNA is easier to isolate than DNA

The mRNA is then combined with free nucleotides and reverse transcriptase to make DNA

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