MadSci Network: Molecular Biology
Query:

Subject: Can one plasmid code for both firefly luciferase and luciferin?

Date: Mon Dec 6 20:42:04 2004
Posted by Scott Heydinger
Grade level: teacher/prof School: Lexington Children's Museum
City: Lexington State/Province: KY Country: USA
Area of science: Molecular Biology
ID: 1102387324.Mb
Message:

GFP bacterial gene transformations are great ways to introduce young kids to 
the relationship between genes and traits.  Unfortunately, most kids in my 
state have never seen fluorescing jellyfish. But they’ve all seen fireflies.

Ward Scientific’s and Promega’s pGL2 firefly based plasmids both code only for 
luciferase and require lysis of the bacteria to release the luciferase and 
addition of luciferin to make the germs produce light.  This is unfortunate, 
since young kids will argue that you don’t need to add chemicals to a 
fireflies’ butt to make it light up.  In contrast, GFP protocols require 
simply adding genes to germs which then “just light up”.

Carolina’s pVIB plasmid (aka pJE202 in Engebrecht) from V. fischeri, also an 
unfamiliar species, contains the lux genes that code for the luciferase enzyme 
and don’t require addition of a substrate, thereby presumably encoding for 
luceferin as well.  No lysis or addition of luciferin are required.

Given my desire to utilize a plasmid based on a familiar species, is it 
possible to have a firefly plasmid similar to pVIB that codes for both 
luciferase and luciferin?  Is such a plasmid available?

Reference:
JoAnne Engebrecht, Kenneth Nealson, and Michael Silverman, “Bacterial 
Bioluminescence: Isolation and Genetic Analysis of Functions from Vibrio 
fischeri,” Cell, Vol. 32, page 773-781, March, 1983.



Re: Can one plasmid code for both firefly luciferase and luciferin?

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