Transcript Biotechnology - BlackSage.com

Biotechnology
Introduction
 Tools
 Process
 Applications
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Biotechnology

Introduction – basic idea
– Gene is identified and excised from one
organism
– Gene is placed in vector (plasmid) and
amplified
– Gene is transferred to new organism or
used in host organism to make protein
product
Biotechnology - Tools
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Restriction endonucleases
– Nucleases cut nucleic acid – at first seem
non specific
– Linn & Abner discover that some strains of
bacteria are able to resist bacteriophage
infection by digesting infecting DNA
– Different bacteria produce different
restriction enzymes
Biotechnology - Tools
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Restriction Endonucleases
– Cut at specific 4, 6, or 8 base sites
– Site is a “palindrome”
» Racecar
» Madam I’m Adam
» Damit I’m Mad
– Some restriction enzymes generate “sticky
ends”
Biotechnology - Tools
Biotechnology - Tools

Plasmids
– Carry an antibiotic resistance marker
– Carry restriction sites in convenient
locations to insert DNA
– Carry characteristics that allow the
plasmid to reproduce in several organisms
Biotechnology - Tools
Biotechnology - Tools
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Polymerase Chain Reaction (PCR)
– Allows any segment of DNA to be
amplified chemically in minutes
– Uses a thermostable DNA polymerase
– Machine can cycle every 60-90 seconds
Biotechnology - Tools
Biotechnology - Tools
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Agarose Gel Electrophoresis
– Can separate DNA fragments made with
restriction enzymes
– Can separate PCR made DNA
– Can be used to sequence DNA
Biotechnology - Tools
Biotechnology - Process
Basic process worked out by Cohen &
Boyer
 Cut plasmid DNA and target DNA
with same restriction enzyme
 Mix DNA and allow sticky ends to
match up
 Select DNA clones having target gene
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Biotechnology - Tools