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Where do mutations come from?
- 1. Biological
- 2. Chemical
- 3. Radiation
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What are biological mutations?
- 1. Mistake in replication
- 2. Recombination
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What are chemical mutations?
- 1. Base analogs
- -stop replication
- -stop transcription
- 2. Intercalating agents
- -Ethidium bromide: fits inside DNA
- 3. Alter DNA
- -Methylate add-CH3-Deaminate
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What are radiation mutations?
- 1. non-ionizing radiation
- 2. ionizing
- -causes electrons to move --> cell damage (proteins, DNA)
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How can you tell a mutation has occurred?
- 1. Cell death
- 2. Cell transformation -->Cancer cell
- 3. Altered metabolism
- 4. Other problems
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How do you assess a chemical? Is it dangerous?
- Rabbit eyes, mice...human
- Use bacteria!
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What are the 3 types of operons?
- 1. Constituitive
- 2. Inducible
- 3. Repressible
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What is a constituitive operon?
Genes always turned "on"
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What is a inducible operon?
Usually off, can turn "on" (lac operon)
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What is a repressible operon?
Usually on, can turn off
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What is Expression?
- Transcription + translation linked
- DNA --> RNA --> Protein
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What are other Eukaryotic controls?
- 1. Amplify DNA
- 2. Inactivate DNA
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What are the tools for genetic manipulation?
- 1. Extract DNA
- 2. Assay (measure)
- 3. Hybridize: to tell similarity
- 4. Cut up DNA
- 5. Separate pieces of DNA
- 6. PCR
- 7. Fingerprints
- 8. Sequence
- 9. Vectors
- 10. Knock out genes
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Explain how to extract DNA
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Explain how to assay
- 1. 260 wavelength in spectrophotometer
- 2. Radioactivity
- 3. Fluorescence
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Explain how to cut up DNA
- 1. Need "scissors"
- -restriction endonucleases
- -restriction enzymes
- -endonucleases
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How do you separate pieces of DNA?
Gel electrophoresis
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What is PCR?
- Polymerase chain reaction
- -gets lots of DNA from small amount
-
Who came up with Taq polymerase?
Brock (the guy who wrote our book!)
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Explain fingerprinting.
Use sections of DNA to identify source
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Explain sequence
Determines order of bases in DNA
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List some Natural vectors.
- 1. transformation
- 2. transduction
- 3. conjugation
- 4. plasmids
- 5. yeast artificial chromosome
- 6. virus
-
List some Artificial vectors.
- 1. DNA bullets
- 2. Microinject
- 3. Electroporation
- 4. Incubate cells in sextrans/CaPO4
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Explain knock out genes.
Take out gene/defective gene
-
What are the states of sewage treatment?
- Prescreen
- 1. Settling
- 2. Bacteria digest
- 3. Advanced, chem, filters
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What are the measurements of viruses?
Most are 20-30 nanometers = .02-.3 micrometers
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Lytic infection
- 1. BO attaches
- 2. Injects DNA
- 3. Bacterium becomes phagefactory
- 4. Phage assemble
- 5. Phage released
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Lysogenic infection
- 1. Attach
- 2. Inject DNA
- 3. Phage DNA incorporated into bacterial chromosome so host has new genes
- 4. Sometimes alter heat/cold other shock
-
+SSRNA
acts as mRNA so host uses to make viral protein
-
-
1934
- FDR birthday balls
- march of dimes
-
1949 Enders, weller and robbin
grow virus in culture, now research really moved fast
-
List +SSRNA
- 1. Picornavirus: cold virus, hepatitis A, polio
- 2. Toga viruses: yellow fever, rubella (German measels)
-
Walter Reed
Army doc, typhoid fever, gets credit for yellow fever
-
-SSRNA
RNA dependent RNA polymerase
-
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Gel electrophoresis
used to separate pieces of DNA
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Restriction enzymes
used to cut DNA
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Hybridization
heating and cooling is used to determine if 2 DNA segments are related
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DNA fingerprints
used to determine if two persons are related
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Polymerase chain reaction
used to make many copies of a DNA segment
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Sequencing
used to determine order of bases in DNA
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Bacteriophage attaches to bacteria
lytic and lysogenic
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Bacteriophage injects its DNA into bacteria
lytic lysogenic
-
host cell begins making viral proteins and viral genetic material, for viral assembly
lytic
-
viral DNA is incorporated into host genome
lysogenic
-
bacterial cell lyses
lytic
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