Midterm 2 Flashcards
1
Q
Antibiotics against bacterial transcription/translation
A
- rifamycin: binds to RNA polymerase
- streptomycin: inhibits peptide initiation and elongation
2
Q
Inducible Operon
A
- turned on by substrate
- usually off
- catabolic operons: breaks down nutrients (lac operon)
3
Q
Repressible Operon
A
- turned off by product synthesized
- usually on
- anabolic operons: enzymes required to synthesize an amino acid stop when no longer needed to be produced (arg operon)
4
Q
Structure of Operon
A
- regulator: gene encoding for repressor protein (on/off switch)
- control locus: promoter locus (binds RNA polymerase) and operator locus (binds the repressor)
- structural locus: actual gene transcribed
5
Q
lac operon
A
- lactose metabolism in e.coli (not e.coli’s first nutrient - glucose is first)
- can be induced on a spectrum
- lactose concentration removes repressor and allows transcription
6
Q
arg operon
A
- regulates arginine synthesis
- excess nutrient binds to repressor and activates it
7
Q
Causes and effects of mutations
A
- causes: spontaneous (random changes arising from error - fluctuation test) or induced (exposure to known mutagens - ames test)
- effects (positive or negative) - mutations are permanent, mainly harmful - can still provide adaptive advantages like antibacterial resistance
8
Q
Ames test
A
- screens mutagenic/carcinogenic compounds
- organism: salmonella typhimurium mutant -> lost the ability to synthesize histidine. (mutation is susceptible to back mutation)
- mutant plated on media with minimal histidine (one sample has mutagen and the other is normal)
- high number of revertant (reverts back to natural) means exposed chemical is mutagenic
9
Q
Fluctuation test
A
- tests for randomly occuring mutation (mutations don’t need to occur by selective agent)
- organism: streptomycin-sensitive organisms
- one sample is cultured in large flask of medium and another is inoculated in 100 tubes of medium
- large flask ~ approximately same number of restant colonies on each plane (always mixing allows even spread of antibiotic resistance)
- inoculated 100 tubes: number of resistant colonies varies widely from plate to plate
10
Q
Horizontal Gene Transfer
A
- movement of any genes between organisms
- G/C content: if there is a large genomic island that is different from core genome and is flanked by direct repeats indicate HGT
11
Q
Transformation Experiment - mice
A
- Fredrick Griffith (1928)
- organism - streptococcus pneumoniae
- four mice injected with rough strain (nonvirulent), smooth strain (virulent), heat-killed smooth strain, and rough strain inoculated with heat killed smooth strain
- shows that bacteria can take up something to become virulent
12
Q
Transformation Experiment - bacteria
A
- Avery & McCarthy (1941)
- Demonstration that DNA is genetic material
- s. pneumoniae is naturally competent (which is rare) - able to take up DNA from environment
13
Q
Gram-positive DNA transformation
A
- dsDNA enters transformasome (DNA binding protein + pore)
- DNA is sucked inside and nuclease makes it ssDNA
- ssDNA is taken by RecA for homologous recombination
14
Q
Gram-negative DNA transformation
A
- pilus binds to dsDNA
- dsDNA becomes ssDNA in periplasm
- other steps are similar to gram positive transformation (RecA recruits ssDNA to homologous site)
15
Q
Bacteria competency
A
- naturally competent
- artificially: induced by electroporation or calcium chloride
- acquired: reponse to signals