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Bio 219 final exam Questions With Complete Solutions Describe the lux operon. What is it, what is its size, wha components does it contain, and what are these individual components responsible for? The lux operon is found in A.fischeri and is responsible for bioluminescence- The operon conta...

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  • November 12, 2024
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  • 2024/2025
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Bio 219 final exam Questions With Complete
Solutions

Describe the lux operon. What is it, what is its size, wha components does it contain, and what

are these individual components responsible for? The lux operon is found in A.fischeri

and is responsible for bioluminescence- The operon contains 5 genes:

luxI- encodes for the auto-inducer

luxR- encodes for the transcriptional activator that binds the auto inducer

luxC- encodes for the acyl-reductase enzyme

luxD- encodes for the acyl-transferase enzyme

luxA- encodes for the alpha subunit of luciferase- has the active site for the enzyme

luxB- encodes for the beta subunit of luciferase- absolutely necessary for the activation of

enzyme

luxE- encodes for the acyl-protein synthetase enzyme




What volume do we run in the reaction? Examples:


If concentration of chDNA is 58 µg/mL = 0.058 µg/µL

Volume of chDNA needed to get us 1.5 µg is:

1.5 µg ÷ 0.058 µg/µL = 25.8 µL of stock chDNA

,Bio 219 final exam Questions With Complete
Solutions
If concentration of vector is 0.2 µg/µL

Volume needed to get us 1 µg is:

1 µg ÷ 0.2 µg/µL = 5 µL of vector stock




If enzyme is at 10 Units/µL

Volume needed to give us 10 U is:

10 U ÷ 10 U/µL = 1 µL of enzyme stock




a 0.7% agarose gel Will show a good separation of larger DNA fragments (5-10 kb)




a 2% agarose gel Will show good resolution for smaller DNA fragments (0.2-1 kb)




Example of a good restriction digestion gel with complete digestion of chDNA and vector

plasmid Lane 1 = undigested


lane 2= partial digestion

lane 3 = complete digestion

, Bio 219 final exam Questions With Complete
Solutions
There are two purposes to our quarter-long project. What are they? a. To create a complete

Aliivibrio fischeri genomic library that will include making E. Coli glow by transformation with

the lux operon

b. To quantify gene expression changes in the transformed E.Coli using gadA




What bioluminescent bacterial organism did we use? Aliivibrio fischeri




Explain the symbiotic relationship between this organism and the creatures that house it.

Symbiotic relationship with a diverse array of marine organisms. Its chromosomal DNA

(chDNA) contains the lux operon which codes for proteins that are necessary for

bioluminescence. Benefits include attracting prey, communication, and hiding from predators.

Benefits to bacteria is reliable food source (glucose and amino acids)




Give two examples of marine organisms that take part in this symbiotic relationship.

Lophius piscatorius (angler fish) and Euprymna scolopes (Hawaiian bobtail squid) (hosts

A. fischeri to hide from predators)




What biochemical reaction causes bioluminescence? Be able to describe and write out the

reaction. FMNH2 + O2 + R-CHO -> FMN + R-COOH +H2O + Light

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