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BiotechnologyBiotechnology
Introduction Tools Process Applications
BiotechnologyBiotechnology
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 - ToolsBiotechnology - Tools
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 - ToolsBiotechnology - Tools
Restriction Endonucleases– Cut at specific 4, 6, or 8 base sites– Site is a “palindrome”
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– Some restriction enzymes generate “sticky ends”
Biotechnology - ToolsBiotechnology - Tools
Biotechnology - ToolsBiotechnology - 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 - ToolsBiotechnology - Tools
Biotechnology - ToolsBiotechnology - Tools
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 - ToolsBiotechnology - Tools
Biotechnology - ToolsBiotechnology - Tools
Agarose Gel Electrophoresis– Can separate DNA fragments made
with restriction enzymes– Can separate PCR made DNA– Can be used to sequence DNA
Biotechnology - ToolsBiotechnology - Tools
Biotechnology - ProcessBiotechnology - 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
Biotechnology - ToolsBiotechnology - Tools