Drexel Bio 219 Final Questions
And Answers Latest Update
Purpose of the Project Answer: 1. to create a complete Aliivibrio fischeri
genomic library that will include making E. coli glow by transformation with the
lux operon
2. create a restriction map of a lux containing plasmid
Aliivibrio fischeri (A. fischeri) Answer: Gram negative, rod shaped bacteria
predominantly found living in symbiosis with a diverse array of marine organisms.
It's chromosomal DNA (chDNA) contains the lux operon, which codes for proteins
that are necessary for bioluminescence
Operon Answer: genetic regulatory system in which genes coding for
functionally related proteins are clustered along the DNA. This feature allows
protein synthesis to be controlled coordinately in response to the needs of the
cell
Chromosomal DNA Answer: large circular piece of DNA (in bacteria). For our
project the chromosomal DNA comes from A. fischeri and we will be isolating the
lux operon from this
,Restriction Digestion Answer: enzymatic reactions that cut DNA into smaller
pieces. Mediated by enzymes that will digest the DNA at only specific sequences
Benefits of bioluminescence to fish Answer: Communication
Attracting Prey
Hiding from predators
Attracting a Mate
Benefits of bioluminescence to bacteria Answer: Reliable food source:
Glucose
Amino Acids
Quorum Sensing Answer: regulation of gene expression in response to
changes in cell- population density
Auto-induction Answer: Bacteria release a small metabolic product, N-Acyl
homoserine lactone, which freely diffuses across the cell membrane and into the
extracellular environment.
As the density of bacteria increases, more of the inducer is released into the
extracellular environment.
When a critical concentration of the inducer is present in the extracellular
environment, the inducer diffuses back into the cells and interacts with the lux
operon - results in gene expression.
, Luciferase Answer: enzyme that catalyzes the light emitting reaction
FMNH2 + O2 + R-CHO FMN + R-COOH + H2O + LIGHT
luxI Answer: encodes for the auto-inducer
luxR Answer: encodes for the transcriptional activator that binds the auto
inducer
luxC Answer: encodes for the acyl-reductase enzyme
Acyl-reductase Answer: Reduces the activated fatty acid to form the
necessary aldehyde
NADPH dependent
luxD Answer: encodes for the acyl-transferase enzyme
Acyl-Transferase Answer: removes fatty acids from their normal biosynthetic
pathway for use in this reaction
luxA Answer: encodes for the alpha subunit of luciferase - has the active site
for enzyme
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