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BiotechnologyThe use of biological processes,
organisms, or systems to manufacture products intended to improve the quality of human life.
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Transgenics• Genetic engineering
makes it possible to transfer DNA sequences from one organism to another
• REMEMBER- DNA is universal
• A gene taken from one species can be replicated in another species.
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Recombinant DNA• Recombinant DNA contains DNA from
2 or more different sources.• So how does DNA get from one species
to another?• Vector: a mechanism needed to transfer
DNA from one organism to another– Microinjection– Gene Gun– Bacteria Plasmid– Virus
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Recombinant DNA• The host cell is often a bacteria cell• Bacteria are used because they have
plasmids• A plasmid is a circular
piece of DNA that exists
apart from the
chromosome and
replicates independently of it.
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• So how do I take a gene from one organism and put it into another?
• We need to use Restriction Enzymes
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What are Restriction Enzymes?
• Proteins• Used by bacteria to cut viral DNA• They “restrict” the growth of viruses
• What does this have to do with transgenics?!
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Restriction Enzymes• Cut strands of DNA at specific nucleotide
sequences• There are many different restriction
enzymes that each cut DNA at different nucleotide sequences
• Usually occurs at a palindrome
5’ GAATTC3’ CTTAAG
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Sticky Ends• Most restriction enzymes cut
DNA with a staggered cut• The staggered cuts leave
the DNA with end pieces “sticking off” – We call these “sticky ends”– These exposed N-bases will
want to join with other complimentary exposed bases
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So what ?• What do you predict could happen if two
pieces of DNA are cut with the same restriction enzyme???– YES! They will have the same “sticky ends”
– How could we use this???
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• Restriction Enzymes can be used to make Recombinant DNA.
• The gene of interest can be isolated using Restriction Enzymes
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Making Recombinant DNA…
• Once the gene is isolated, how do we join it with the organism’s DNA?
• Cut the organism’s DNA with the same restriction enzyme…why– The sticky ends will naturally be attracted to
each other• Add DNA LIGASE: an enzyme that seals
the fragments together
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• Now whenever the bacteria plasmid replicates, the gene will be cloned.
• Gene Cloning: the production of many identical copies of the same gene.
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• You have created a Transgenic Organism!
–organisms that contain functional recombinant DNA (rDNA) from a
different organism
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• http://www.sumanasinc.com/webcontent/animations/content/plasmidcloning.html