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Where do mutations come from?
- 1. Biological
- 2. Chemical
- 3. Radiation
What are biological mutations?
- 1. Mistake in replication
- 2. Recombination
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
What are radiation mutations?
- 1. non-ionizing radiation
- 2. ionizing
- -causes electrons to move --> cell damage (proteins, DNA)
How can you tell a mutation has occurred?
- 1. Cell death
- 2. Cell transformation -->Cancer cell
- 3. Altered metabolism
- 4. Other problems
How do you assess a chemical? Is it dangerous?
- Rabbit eyes, mice...human
- Use bacteria!
What are the 3 types of operons?
- 1. Constituitive
- 2. Inducible
- 3. Repressible
What is a constituitive operon?
Genes always turned "on"
What is a inducible operon?
Usually off, can turn "on" (lac operon)
What is a repressible operon?
Usually on, can turn off
What is Expression?
- Transcription + translation linked
- DNA --> RNA --> Protein
What are other Eukaryotic controls?
- 1. Amplify DNA
- 2. Inactivate DNA
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
Explain how to extract DNA
Explain how to assay
- 1. 260 wavelength in spectrophotometer
- 2. Radioactivity
- 3. Fluorescence
Explain how to cut up DNA
- 1. Need "scissors"
- -restriction endonucleases
- -restriction enzymes
How do you separate pieces of DNA?
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!)
Use sections of DNA to identify source
Determines order of bases in DNA
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
Explain knock out genes.
Take out gene/defective gene
What are the states of sewage treatment?
- 1. Settling
- 2. Bacteria digest
- 3. Advanced, chem, filters
What are the measurements of viruses?
Most are 20-30 nanometers = .02-.3 micrometers
- 1. BO attaches
- 2. Injects DNA
- 3. Bacterium becomes phagefactory
- 4. Phage assemble
- 5. Phage released
- 1. Attach
- 2. Inject DNA
- 3. Phage DNA incorporated into bacterial chromosome so host has new genes
- 4. Sometimes alter heat/cold other shock
acts as mRNA so host uses to make viral protein
- FDR birthday balls
- march of dimes
1949 Enders, weller and robbin
grow virus in culture, now research really moved fast
- 1. Picornavirus: cold virus, hepatitis A, polio
- 2. Toga viruses: yellow fever, rubella (German measels)
Army doc, typhoid fever, gets credit for yellow fever
RNA dependent RNA polymerase
used to separate pieces of DNA
used to cut DNA
heating and cooling is used to determine if 2 DNA segments are related
used to determine if two persons are related
Polymerase chain reaction
used to make many copies of a DNA segment
used to determine order of bases in DNA
Bacteriophage attaches to bacteria
lytic and lysogenic
Bacteriophage injects its DNA into bacteria
host cell begins making viral proteins and viral genetic material, for viral assembly
viral DNA is incorporated into host genome
bacterial cell lyses