Process for producing a cloned luciferase-synthesizing microorganism
AssigneeThe Texas A&M University System
InventorThomas O. Baldwin
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Abstract
A process for producing a cloned luciferase-synthesizing microorganism, capable of the expression of bioluminescence, is disclosed. The process involves isolation and digestion of the genomic DNA of an appropriate bioluminescent microorganism, such as Vibrio harveyi, to obtain a DNA fragment which codes for luciferase. The DNA fragment is inserted into a pre-existing cloning vehicle under the control of an expression promoter, thereby producing a recombinant or derivatized cloning vehicle. A host microorganism is transformed with the derivatized cloning vehicle, cultured and purified for the expression of bioluminescence.
Claims16
A process for producing a cloned luciferase-synthesizing microorganism capable of expressing bioluminescence comprising:
- isolating genomic DNA of Vibrio harveyi;
- digesting said genomic DNA to obtain a DNA fragment comprising an approximate 4.0 kilobase pair restriction fragment encoding for both the alpha and beta subunits of luciferase-synthesizing gene;
- inserting said DNA fragment into a DNA cloning vehicle able to replicate in a host microorganism proximate to a promoter thereby producing a derivatized cloning vehicle;
- transforming a host microorganism with said derivatized cloning vehicle to produce a transformed microorganism which expresses bioluminescence;
- culturing said transformed microorganism;
- and
- purifying said transformed microorganism for the expression of bioluminescence.
A cloning vehicle having a DNA sequence which is able to replicate in a host microorganism, said cloning vehicle comprising:
- a DNA fragment from Vibrio harveyi comprising approximately 4.0 kilobase pair restriction fragment encoding for both the alpha and beta subunits of luciferase-synthesizing gene;
- and
- an expression promoter in operable proximity to said DNA fragment.
A transformed microorganism E. coli RR1/pTB7.
Dependencies are inferred from the claim text.