Biotechnology You Will Learn About… Transformation Cloning DNA Fingerprinting by Restriction Fragment Length Polymorphism (RFLP) What is the name of the lamb?

Brainstorm What are some ways in which DNA can be used as a tool (think about TV, the news…)

Goals of Applied Genetics 1.Help humans create crops that can be frost resistant (we love our peaches!) 2.Use transgenic organisms to help medical researchers model human physiology for testing 3.Help industry to create bacteria to break down pollutants into harmless products 4.Pharmaceutical companies use recombinant DNA to cheaply produce human hormones (insulin) and other proteins 5.Help solve crimes and determine relatives

Technologies Addressed in this Show please write these on first slide –Bacterial Transformation –Cloning –DNA fingerprinting (RFLP analysis)

Selective breeding Selecting organisms with the most desirable traits Requires time and several generations to produce offspring with the desired trait Ex: –Short vs long haired cats –Milk production in cattle –Disease resistant foods –Bacteria that break down oil

Bacterial Transformation Also called recombinant DNA technology or gene cloning Uses a bacterial host because of fast reproduction and a circular DNA vehicle to hold the foreign DNA=plasmid Organisms containing recombinant DNA or foreign DNA are known as transgenic

Steps involved: 1.Select the desired gene(s) to be inserted into the organism and a bacterial host containing a plasmid (vehicle to hold the desired gene) 2.Cut two DNA molecules into fragments with special (restriction) enzymes 3.Splice (rejoin) the fragments (gene and plasmid) together in the desired combination 4.Introduce or insert the new DNA into a living cell for replication (mitosis)

= Vector to transfer DNA

Enzymes Restriction Enzymes Used to cut a DNA molecule at a specific nucleotide sequence Produces one of two types of DNA fragments 1.Sticky ends (palindrome) 2.Blunt ends

Sticky Ends (palindrome) Blunt ends

Gene Cloning Used to produce genetically identical copies of a cell, tissue, organ, and/or organism Needed to produce multiple copies of the desired DNA

Cloning Applications Currently Plants are cloned to produce a large number of genetically identical plants in a short amount of time Future 2.Clone productive and healthy animals to increase yield for farmers and to grow organs for transplants

How cloning works….

Gene Therapy The insertion of normal genes into human cells to correct genetic disorders like cystic fibrosis. (Not successful yet?)

DNA Fingerprinting (RFLP Analysis) 1.Obtain a small sample of DNA 2.Make millions of copies using PCR (polymerase chain reaction) technique 3.Cleave (cut) DNA with restriction enzymes 4.Separate DNA fragments using gel electrophoresis and compare 5.Each human’s DNA will have some unique pieces because each of our DNA is unique

Gel Electrophoresis Gel electrophoresis is used to create a DNA fingerprint Small amounts of DNA are loaded into wells in the gel. An electric current pushes the small pieces of DNA farther down the gel than the larger pieces. DNA patterns are compared to known patterns (Human Genome Project). Used for forensics, blood samples, and paternity tests (children get half their bands from mom and half from dad)

Running a gel…

How DNA moves… Step 1: Make wells in gel.

Step 2: Pipette DNA into wells.

Step 3: Run an electric current to watch DNA migrate.

Step 4: Notice similarities and differences in banding patterns.

The Human Genome Project There are approximately 25,000 genes on the 46 human chromosomes (far fewer than expected) Human Genome Project- an international effort to completely map and sequence human chromosomes (completed April 2003) This has been important in –identifying genes that are related to disease. –Determining human relationships with other species.

Microarray An new technology, the microarray, allows a cell to be samples for many genes at once. It is involved in finding –What genes are important in cancer and cancer prevention. –What medicines can be used for a person (customized medicine) –Much more, and we will spend some time with this.