Biotechnology ppt notes Recombinant DNA Purpose: To ___________of one organism into another organism BSCS Biology/ Juo

Recombinant DNA Applications Biotechnology ppt notes Recombinant DNA Applications Green-glowing aquarium fish (jellyfish genes) Fast-growing fish (Salmon with Pout genes) Herbicide-_____________ crops (Round-Up Ready) http://news.bbc.co.uk/2/hi/science/nature/3026104.stm BSCS Biology/ Juo

_____-resistant crops (__ toxin) Biotechnology ppt notes _____-resistant crops (__ toxin) BSCS Biology/ Juo

Recombinant DNA Applications Biotechnology ppt notes Recombinant DNA Applications _________ production (bacteria can make insulin, hGH, etc) BSCS Biology/ Juo

Making Recombinant Bacteria Important players: ________ of interest (i.e. insulin, hGH, etc) ______________ enzymes – cut the DNA ___________ – circular DNA found in bacteria ________________ BSCS Biology/ Juo

What are Restriction Enzymes? Target very ___________ ______ sequences Are found in more than 100 different varieties Are used in nature to ______________ bacteria from foreign invaders BSCS Biology/ Juo

What are Restriction Enzymes? Biotechnology ppt notes What are Restriction Enzymes? Each restriction enzyme recognizes a very specific nucleotide _______ EcoR1 recognizes: GAATTC CTTAAG The enzyme cuts it: G AATTC CTTAA G BSCS Biology/ Juo

BSCS Biology/ Juo

Biotechnology ppt notes BSCS Biology/ Juo

Making Recombinant DNA Biotechnology ppt notes Making Recombinant DNA Plasmids - self-replicating rings of _______ containing 2-30 genes, found in bacterial cells BSCS Biology/ Juo

Making Recombinant DNA Biotechnology ppt notes Making Recombinant DNA __________ the gene of interest using a restriction enzyme Cut the __________ (using the same enzyme) __________ gene into the plasmid Insert the plasmid into _________ Grow bacteria and harvest the ____________ BSCS Biology/ Juo

Making a paper bracelet You will need: A small circle of paper (this is your plasmid) A strip of paper in a different color about 5 inches long. In the center is a 1-2 inch piece that is your gene of interest. Cut out the gene with your scissors. Cut the circle once and using a glue stick or tape, add your gene to the open circle to form a closed circle.

Biotechnology ppt notes 1. Isolating the gene Use restriction enzymes to cut the DNA strand at specific places that surround the gene BSCS Biology/ Juo

2. Cut the plasmid Cut the plasmid using the __________ restriction enzyme BSCS Biology/ Juo

3. Put the gene in the plasmid Since the gene and the plasmid have the same ______________ ________, the gene will stick to the plasmid BSCS Biology/ Juo

Biotechnology ppt notes The gene of interest is attached to a promoter and some kind of marker BSCS Biology/ Juo

5/6. Put plasmid in bacteria Insert plasmid into bacteria Let bacteria make the protein BSCS Biology/ Juo

Cloning a gene: http://highered.mcgraw-hill.com/sites/0072437316/student_view0/chapter16/animations.html# BSCS Biology/ Juo

Biotechnology ppt notes DNA Fingerprinting Definition: Creating a __________ _______ ______________ for someone Purpose: Forensic science Determining paternity Research Diagnosing Disease BSCS Biology/ Juo

Gel Electrophoresis Analogy (DO NOT COPY THIS DOWN) Biotechnology ppt notes Gel Electrophoresis Analogy (DO NOT COPY THIS DOWN) "Imagine a huge swimming pool full of water and many entangled nets (this is the agarose gel). A dump truck comes along and dumps its load into one end of the pool. That load contains an extensive collection of long, skinny things of varying lengths and sizes (analogous to DNA of varying sizes), e.g. from boa constrictors to smaller snakes to worms to microscopic bacteria. There is a vacuum at the other end of the pool (analogous to the electric current), which can be turned on and off. When on, the vacuum pulls all of the creatures across the pool. But because of the netting, movement of the skinny things is impeded." BSCS Biology/ Juo

Biotechnology ppt notes Gel Electrophoresis Separates DNA fragments by _________ using ______________ current BSCS Biology/ Juo

Biotechnology ppt notes Gel Electrophoresis Larger fragments move more _________ Bands of fragments result BSCS Biology/ Juo

DNA Fingerprinting Method Biotechnology ppt notes DNA Fingerprinting Method Use restriction enzymes to cut the DNA Load the DNA onto agarose gel for gel electrophoresis Analyze the banding pattern BSCS Biology/ Juo

Biotechnology ppt notes BSCS Biology/ Juo

Gel Electrophoresis BSCS Biology/ Juo

Gel Electrophoresis BSCS Biology/ Juo

RFLP Analysis RFLP – Restriction Fragment Length Polymorphism: for related DNA molecules, a difference in DNA fragment sizes after restriction enzyme digestion Difference results from presence of different DNA sequences Certain regions of genome are highly variable BSCS Biology/ Juo

A single nucleotide change can make a difference Wild-type allele AGATCT TCTAGA Restriction site Mutant allele AGAGCT TCTCGA Not a restriction site BSCS Biology/ Juo

Cut site present = 2 shorter DNA fragments No cut site = 1 long DNA fragment RFLP Cut site present = 2 shorter DNA fragments

NOVA: Cracking the Code - #9: Finding Cures is Hard http://www.pbs.org/wgbh/nova/genome/program.html