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Bio 219 Final Exam Review Questions with Correct Answers
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Bio 219 Final Exam Review Questions with Correct Answers
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Bio 219 Final Exam Review Questions with CorrectAnswers
1. Two main goals of project: 1. Isolate the lux operon out of A.fischeri
and use it to transform E.coli to induce bioluminescence
2.Create a restriction map of a lux-containing plasmid (OR detect GadA
expression levels)
2. Bioluminescent bacterial organism used?: A.fischeri
3. Explanation of symbiotic relationship between this organism and hosts-
: A.fischeri gets glucose and amino acids from host, host can camouflage
from predators, attract prey, or attract mates
4. Two marine organisms that take part in this symbiotic relationship:
Hawaiian bobtail squid and Angler fish
5. Lux operon: Definition, Size, Components and their Responsibilities:
Lux operon induces bioluminescence in A. fischeri
8.5kb long
luxI-encodes for auto-inducer
luxR-encodes for transcriptional activator that binds to
auto-inducer luxC-acyl-reductase enzyme
luxD-acyl-transferase enzyme
1
,luxA-alpha subunit of luciferase (has active site)
luxB-beta subunit of luciferase (necessary for activity of
enzyme) luxE-acyl-protein synthetase enzyme
6. Biochemical Rxn that causes Bioluminescence: •FMNH2 + O2 + R-
CHO à FMN + R-COOH + H2O + LIGHT
Luciferase catalyzes light emitting rxn
Acyl-transferase removes fatty acids from normal pathway for use in
rxn, acyl-pro- tein synthetase activates fatty acid to form R-CO-AMP
(use ATP), acyl-reductase reduces activated fatty acid to form
necessary aldehyde (use NADPH)
7. Quorum sensing: Regulation of gene expression in response to
changes in cell-population density
8. Auto-induction: Bacteria release AHLs (N-acyl homoserine lactone) to
freely diffuse into extracellular environment. As bacteria density
increases, more inducer is released. When a certain concentration is
hit, inducer diffuses back into cells to interact with Lux operon (turn on
gene expression)
9. E. coli LuxR homolog?: SdiA
10.Can E.coli use quorum sensing? How do they do this in nature: Can use
2
, quorum sensing from other organisms that produce AHLs as a means to
up or down regulate parts of its genome
11.GadAx operon: Definition, Encodes for, Gene of Focus: Operon that
allows E.Coli to survive stress (overexpressed during times of stress).
Encodes for a glutamate decarboxylase that plays a role in acid
tolerance Gene of Focus: GadA gene
12.What happens to GadAx expression in presence of lux? Reference
gene used for comparison: In presence of lux and the AHL it produces,
GadA is upregulated.
16sRNA
13.Two samples used as comparison on how gene expression changes
in presence of lux operon: GadA and 16SrRNA
14.Structure of bacterial chromosomes: Circular, gram negative have
thin layer of peptidoglycan-based cell wall
15.Reagents needed for lysis phase of chDNA isolation: Resuspension
buffer, Proteinase K, RNaseA, Lysis buffer
16.Resuspension buffer: Breaks down cell membranes (contains SDS)
and allows for pellet resuspension
17.Proteinase K: Breaks down proteins and nucleases
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