Thermofluor

Summary of Fluorescent Protein Gene Products at Lybradyn

As part of Lybradyn's protein expression product and technology research a series of genes have been identified from a novel species of the extreme thermophile geobacillus. Two genes are responsible for the fluorescence observed one imparting an emission in the red and the other in the green portions of the visible spectrum. Both proteins show excitation maxima in the long ultraviolet region. Both genes have been cloned, sequenced and over expressed in e coli.

The gene coding for the protein responsible for the red fluorescence has been identified as a uroporphyrinogen methyltransferase and functions as a key step in porphyrin biosynthesis. Over expression of the gene produces a buildup of fluorescent porphyrin pigments. The gene appears to be stable in both bacillus and e coli systems and can be used as a genetic marker for expression systems requiring visual display of recombinant expression. Neither the pigment nor the protein appears to create acute toxicity issues for host cells.

The gene has been studied in a number of bacterial systems and has even been proposed as a genetic marker in the past (ref). In our hands we are currently developing expression vectors that contain this gene as a indicating marker for cloning, genetic manipulation and expression optimization.

The second gene that we are working with displaying fluorescent properties is from the same thermophilic organism. The gene has been cloned, sequenced and over expressed in e coli and bacillus and has been identified as coding for hihydroorotate dehydrogenase. The protein is a flavoprotein and is a key to pyrimidine biosynthesis. The enzyme, much like GFP displays intense green fluorescence when excited with long ultraviolet light. We have used this gene as a marker for expression and is being considered for potential for fusion protein applications.

For both add Molecular weight and other tech data like the spectra.