Gene Technology

What Is Gene Technology? Gene Tech  the alteration of genes inside a living organism to produce a specific results Examples? Examples? GM Foods: GM Foods: Disease resistant potatoes Disease resistant potatoes Pesticide producing tomatoes Pesticide producing tomatoes Golden Rice Golden Rice GM Pets: GM Pets: Cloned pets Cloned pets Glow in the dark pets Glow in the dark pets

Gene Technology Vocab Crunch! Target gene  DNA we want to work with Target gene  DNA we want to work with Reverse transcriptase  enzyme that makes DNA from mRNA Reverse transcriptase  enzyme that makes DNA from mRNA cDNA  complementary DNA strands made from mRNA cDNA  complementary DNA strands made from mRNA DNA Polymerase  copies DNA DNA Polymerase  copies DNA Vector  object DNA will be placed into Vector  object DNA will be placed into Plasmid  small ring of DNA that can be added to a bacterial genome Plasmid  small ring of DNA that can be added to a bacterial genome DNA Ligase  reconnects sections of DNA backbone (C-P-C) DNA Ligase  reconnects sections of DNA backbone (C-P-C) Restriction Enzymes  cut DNA and plasmid in specific places so to make “sticky ends” (matching ends) Restriction Enzymes  cut DNA and plasmid in specific places so to make “sticky ends” (matching ends)

I totally didn’t steal this image

Steps of Producing Insulin 1)mRNA is extracted from pancreas cells 2)Reverse transcription using reverse transcriptase 3)Single stranded DNA is made (cDNA) 4)DNA Polymerase is used to make dsDNA 5)Restriction enzyme is used to cut dsDNA to make sticky ends 6)Same restriction enzyme is used to cut plasmid vector

Steps of Producing Insulin (II) 7) Open plasmid vector, ds DNA, and DNA ligase all mixed together 8)Plasmid vector produced with insulin gene inside (WOOT!) 9)Plasmid vector added to bacteria by transformation (electric shock) 10) Grow bacteria and test for insulin gene 11) Grow bacteria that successfully make insulin 12) Isolate, clean, and sell insulin protein

*le badly drawn mRNA Insulin mRNA isolated RT *le incredibly well drawn cDNA

DNA polymerase *le ok-drawn dsDNA Restriction Enzyme dsDNA with sticky ends

AATT TTAA dsDNA with sticky ends

Same Restriction Enzyme AATT TTAA Plasmid Cut plasmid

AATT TTAA AATT TTAA DNA Ligase

Insulin gene Bacteria Transformation

Antibiotic selection PCR testing for gene

How Do We Know It Worked? 1)Antibody selection  Add in penicillin resistant gene to plasmid; only bacteria with plasmid should grow Add in penicillin resistant gene to plasmid; only bacteria with plasmid should grow 2)PCR sample and run on a gel  Make copies of bacteria plasmid and digest with same restriction enzymes and separate on a gel Make copies of bacteria plasmid and digest with same restriction enzymes and separate on a gel Same band = same gene Same band = same gene 3)Green Fluorescent protein  Add in gene to make green fluorescent protein Add in gene to make green fluorescent protein Only green bacteria have plasmid Only green bacteria have plasmid

Verbal cards Debate: “Should there be more regulation on the production and distribution of GM foods?”