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Chapter 16 and 13.


Genetic code is universal
 Translation results in the same amino acid sequence regardless of organism

Opens the doors to techniques to analyze and manipulate DNA
 Cutting and transferring genes from one organism to another  Copying DNA in a lab  Using DNA to reveal identity  Mapping nucleotide sequence  Cloning

each cuts at a different sequence  Ex.coli)  Protects cells from invading DNA .   Hybrid molecule composed of DNA from 2 or more organisms Requires restriction enzymes and DNA ligase Restriction enzymes cut DNA at a specific nucleotide sequence  Various restriction enzymes.EcoRI (found in E.

Enzymes recognize short nucleotide sequence (restriction site)  Symmetrical.1.sequence is the same on both strands when read in the 5’  3’ direction 2. Cleave backbonesingle stranded end = sticky end .

Sticky ends will temp bond with complementary sticky ends of other DNA  Source of DNA doesn’t matter  Use the same restriction enzyme for all the sources of DNA 4.3. DNA ligase seals the backbone .

  Recombinant DNA is first step in DNA cloning Allows researchers to generate clones of bacterial cells with the foreign gene  Use plasmids. small circular pieces of DNA in bacteria (not part of main chromosome)  Foreign DNA and plasmid treated with same restriction enzyme and mixed together  Sticky ends join  Modified plasmid reintroduced into cell (transformation)  Just before division cell DNA and plasmids replicate  Daughter cells both have the modified plasmid  Researchers harvest DNA from clones .

heat stable polymerase.  Cloning in cells is best for preparing large quantities If DNA source is scanty or impure. is quicker and more selective  Ex. polymerase chain reaction (PCR).forensics   Requirements: DNA. primers Temperature (heat) is used instead of helicase . free nucleotides.

heat stable DNA polymerase extends primers in 5’  3’ direction  Left to run for several hours and will complete approx 20 cycles  End result = copy number is ~1 million times original number PCR animation . 3 step cycle  Denaturation.heated and strands separate  Annealing. allows primers to attach  Extension.strands cooled.

take the quiz and email me your results to dodi.  Click on the following links and look at the This must be done by next rotation.  DNA Cloning  PCR . Once it is complete.fl.

    Process which uses a gel medium to separate nucleic acids fragments according to size DNA samples are “loaded” into the gel Gel is bathed in an aqueous solution and has electrodes attached to each end Current is turned on  Fragment separation is based on charge and size  Small fragments will move faster and further away from starting point  DNA will move toward which electrode?  positive  DNA is dyed and then analyzed .

 Useful for separating DNA fragments produced by restriction enzymes  Prepare pure samples of individual fragments   Compare 2 different DNA molecules Can be used to sequence a DNA fragment Lab simulation/ HW:  Electrophoresis  Wkst on edline  .

individuals are related . DNA has repeating sequences in noncoding regions     Known as tandem repeats Multiple copies of a 2-10 base sequence High rate of mutation at these locations Different individuals have a unique length to these regions    PCR copies a region of DNA known to have tandem repeats Gel electrophoresis is used and the resulting bands constitute an individual’s DNA fingerprint Banding pattern of DNA sample is matched with known samples  If identical. they came from same individual  If similar.

  Chance of 2 people (not identical twins) having identical tandem repeats in 3 regions is 1:1018 Uses:  Forensics (blood. hair follicle) figure 16. semen.child’s DNA is a combination of both parents.9 on p 247  Paternity suits. bands from a child must come from either mom or dad  Study population dispersals of humans or other animals  Trace individual’s ethnic heritage .

copy it.  Started in .000 genes in human DNA   Now its deciphering what each gene does Generated genome libraries of genetic diseases  Aids doctors in diagnosing  New medication production  Where gene is.ornl. synthesize molecule and distribute   Ancestry and migration patterns http://www. completed in 2003 Goal was to determine the order of all bases in human DNA  Approx 30.

 Involves deliberate changes to an individual’s genome  Usually through a plasmid or virus  Results in genetically modified organisms (GMO’s)  Express the transplanted gene and synthesize proteins  Bacteria and yeast most common GMO’s  Insulin.oil spill)  Food manufacturing . and tissue plasminogen activator (TPA) currently produced this way  Organisms for toxic chemical cleanup (ex. growth hormone (hGH).

  Genetic engineering has been used for years in plants Examples  Delayed ripening (off season veggies)  Resistance to spoilage or drought  Resistance to the herbicide glyphosate (akaRoundUp)  Improve nutritional value-”golden” ricecontains beta carotene (makes vit A.trials for Hep B vaccines.for eyesight and proper brain development)  “Pharm” plants. antibodies that interfere w/ the bacteria that cause tooth decay .

pig w/ leaner meat “Pharm” animals. heart attacks. blood clotting disorders. Mice were first transgenic animals  Allows us to study human diseases w/o experimenting on humans   Ex.sheep w/ better wool.produce biological substances for medical use  Pigs potential source for transplant organs  Goats produce proteins to treat cystic fibrosis. etc  Sheep altered to secrete Factor IX protein in their milk  Treats hemophilia  Some vaccines produced in chicken eggs  Be careful! Ensure no allergic or other adverse reactions to patients receiving them. (egg allergy) .

 Good     Improve production Better environment Reduce hunger in nations w/ poor crop output Produce rare proteins for vaccines Are GMO (genetically modified organisms) safe to eat? No long term data Transgenic plants could pass new genes to wild crops Products could produce allergic reactions Genes could cross species Decrease in biodiversity  Bad       .

Nucleus removed from one of the cultured cells.  Unfertilized egg from surrogate mom  Using a differentiated animal cell  “Dolly”  Somatic cell from donor sheep collected and cultured. Nucleus removed from egg. .  Egg cell and nucleus from somatic cell fused together w/ a zap of current  Embryo placed in “mom” and developed normally  Baby is a clone of the original donor. http://learn.genetics.utah.

 Using undifferentiated cells  Therapeutic cloning  Referred to as stem cell research  Major ethical issue:  Produces human embryos for science  Benefits:  Repair damaged nervous tissue (spinal cords)  Grow skin for serious burns  Grow heart muscle to repair a damaged heart .