1k likes | 1.01k Views
Biotechnology. Biotechnology. “Biotechnology means any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use.”. Bacteria with Plasmid. 1 = bacteria regular chromosome 2 = plasmids. Plasmid.
E N D
Biotechnology • “Biotechnology means any technological application that uses biological systems, living organisms, or derivatives thereof, to make or modify products or processes for specific use.”
Bacteria with Plasmid 1 = bacteria regular chromosome 2 = plasmids
Plasmid • Small circular DNA separate from the larger bacterial chromosome
Why is a plasmid important in biotechnology? • It may be used as a vector, to carry other genes to another cell.
Gene therapy is being used in many ways. For example, to: • Replace missing or defective genes; • Deliver genes that speed the destruction of cancer cells; • Supply genes that cause cancer cells to revert back to normal cells; • Deliver bacterial or viral genes as a form of vaccination; • Provide genes that promote or impede the growth of new tissue; and; • Deliver genes that stimulate the healing of damaged tissue.
R plasmids • This type of plasmids carry genes for enzymes that destroy antibiotics such as penicillin and tetracycline. • So…Bacteria containing R plasmids become resistant to many antibiotics.
Steps for Gene Therapy • 1. Isolate plasmid
Animation • LINK: Isolating Plasmid DNA from Bacteria
3. Gene is inserted into plasmid RECOMBINANT DNA = plasmid + inserted DNA
4. Recombinant DNA plasmid is put into bacterial cell • DNA cloning animation • 5. Cell multiplies with recombinant DNA
Restriction enzyme • (or restriction endonuclease) is an enzyme that cuts double-stranded DNA at a specific location
Different Restriction Enzymes Several hundred different kinds LINK: RECOGNITION SITES OF RECTRICTION ENZYMES
“Sticky Ends” Complementary bases join plasmid an new DNA to form recombinant DNA Restriction site = place where the two fragments join
Ligase Enzyme Joins the sticky ends
Recombinant DNA • Plasmid DNA + other organism DNA • LINK: rDNA
DO LAB ACTIVITY Chocolate Flavored Cherries: An exercise in Recombinant DNA Technology Read intro and do Pre-Activity First
Gel Electrophoresis • Gel Electrophoresis Virtual Lab • Gel Electrophoresis
Ready to Run • DNA flows from – to + end of the gel
DNA Fingerprinting Each person produces a specific pattern of bands of DNA sequence lengths
Examine the results • UV Transilluminator, and the UV light shines up through the gel.
Click and Drag to find the suspect Murder at Rodman Dam • http://www.dnalc.org/ddnalc/resources/shockwave/dnadetective.html
Some Examples of DNA Uses for Forensic Identification • Identify potential suspects whose DNA may match evidence left at crime scenes • Exonerate persons wrongly accused of crimes • Identify crime and catastrophe victims • Establish paternity and other family relationships
More Uses • Detect bacteria (like Salmonella) and other organisms that may pollute air, water, soil, and food • Match organ donors with recipients in transplant programs • Determine pedigree for seed or livestock breeds
More Uses • Authenticate consumables such as caviar and wine • Detect disorders like cancer, cystic fibrosis • Identify endangered and protected species as an aid to wildlife officials (could be used for prosecuting poachers)
OTHER FRAGMENTS: VNTR’s • Variable number of tandem repeats (VNTRs) can be used in a gel electrophoresis to show different patterns
Southern Blot Animation • LINK: Animation Quiz 5 - Southern Blot
Paternity Testing: Call Montel • NICE LINK Paternity Testing
Would any one else have the same pattern as you? • No…unless you are an identical twin.
DNA PROBES • a single-stranded DNAmolecule (like ATTGGAC) used in laboratory experiments to detect the presence of a specific sequence among a mixture of other singled-stranded DNA molecules.
USES of DNA PROBES • Detect infectious diseases • Detect genetic diseases • HLA typing (suitable donor for stem cell blood typing) • Paternity (find the father) • Cancer detection
PCR • Polymerase Chain Reaction- • take a small sample of DNA an amplify it take make a larger amount • Sample does not have to be pure and can be partially degraded.