Presentation on theme: "Genetic Engineering (and other cool molecular biology techniques)"— Presentation transcript:
Genetic Engineering (and other cool molecular biology techniques)
DNA Cloning DNA cloning permits production of multiple copies of one particular gene This gene copy will be used to produce a usable protein product (e.g. insulin) or for genetic engineering (e.g. gene for pest control inserted into plant)
Transformation Get “competent” bacteria (usually E.coli that is capable of being transformed) Treat those cells to calcium chloride Heat shock to force bacterial cell to take up plasmid DNA
Now… Select for bacteria that have taken up your plasmid (usually use antibiotic) Grow bacteria in large quantities, purify out protein, and…
PCR (polymerase chain reaction) Specific sequence of DNA is amplified (copied many times) Requires: – DNA template (contains your gene of interest) – Tac polymerase (a DNA polymerase that can work at high temperatures) – Nucleotides (to synthesize new DNA) – Primers (specific to the gene of interest)
PCR is a cyclic process: PCR is a cyclic process – Heat to denature (open up) DNA – Cool to allow primers to anneal (stick to) single stranded DNA – Warm to allow DNA to be replicated by polymerase – REPEAT REPEAT
1. Genetically Modified Food Insert particular genes within a type of food to make it better tasting, longer lasting, or more healthy. Long term effects of consumption of this food are unknown.
2. Transgenic Animals Research purposes – Knock outs – Overexpression – Use of GFP to select for animals that are transgenic Consumer purposes (food or medication)
3. Pharmaceuticals Use plants or other GMO to produce pharmaceuticals – Ex: Gaucher’s disease (lysosomal storage disease – buildup of fat in cells); Pfizer received patent to grow drug to treat this disease in tobacco plant – Ex: Use of a transgenic goat to produce an anticoagulant (in the goat milk) to reduce probability of blood clots Use bacteria to produce insulin, human growth hormone, clotting factors for hemophaelics GMO mammals (mice) used in pharmaceutical research labs to study disease and treatment
Your Homework: Read the entire laboratory handout. You must carefully read the background information AND the laboratory procedure as well. Answer the pre-lab questions.