US4581335A
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Process for producing a cloned luciferase-synthesizing microorganism
Assignee: Texas A&M University System
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Filing: 1982-12-01
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Granted: 1986-04-08
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Expires: 2002-12-01
Abstract
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.
Inventors
- 👤 Thomas O. Baldwin
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-30 05:32
CPC Classifications
C; C12; C12N; C12N9/00; C12N9/0004; C12N9/0071; C12Y; C12Y114/00; C12Y114/14; C12Y114/14003; Y; Y10; Y10S; Y10S435/00; Y10S435/8215; Y10S435/822; Y10S435/848; Y10S435/849
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
104 words
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.
Claim 2
Dependent Claim
104 words
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.
Claim 3
Dependent Claim
13 words
The process of claim 1 wherein said DNA cloning vehicle is plasmid DNA.
Claim 4
Dependent Claim
12 words
The process in claim 1 wherein said DNA cloning vehicle is pBR322.
Claim 5
Dependent Claim
12 words
The process in claim 1 wherein said derivatized cloning vehicle is pTB7.
Claim 6
Dependent Claim
18 words
The process in claim 1 wherein said DNA fragment is approximately a 4.0 kilobase pair Hind III fragment.
Claim 7
Dependent Claim
33 words
The process in claim 1 wherein said DNA fragment is approximately a 4.0 kilobase pair fragment selected from one of SalI-EcoRI; SalI-Hind III; SalI-BamHI; BamHI-EcoRI; BamHI-Hind III; BamHI-BamHI; Hind III-EcoRI; or Hind III-BamHI.
Claim 8
Dependent Claim
55 words
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.
Claim 9
Dependent Claim
55 words
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.
Claim 10
Dependent Claim
13 words
The cloning vehicle of claim 7 wherein said DNA sequence is plasmid DNA.
Claim 11
Dependent Claim
19 words
The cloning vehicle of claim 7 wherein said DNA fragment is approximately a 4.0 kilobase pair Hind III fragment.
Claim 12
Dependent Claim
31 words
The cloning vehicle of claim 7 wherein said DNA fragment is approximately a 4.0 kilobase pair selected from SalI-EcoRI; SalI-Hind III; SalI-BamHI; BamHI-EcoRI; BamHI-Hind III; BamHI-BamHI; Hind III-EcoRI; or Hind III-BamHI.
Claim 13
Dependent Claim
14 words
The cloning vehicle of claim 8, wherein said cloning vehicle is derived from pBR322.
Claim 14
Dependent Claim
18 words
The cloning vehicle of claim 11 wherein the promoter is located in the tetracycline resistance region of pBR322.
Claim 15
Dependent Claim
12 words
The cloning vehicle of claim 8, wherein said cloning vehicle is pTB7.
Claim 16
Dependent Claim
17 words
A microorganism transformed by a cloning vehicle of claims 7, 8, 9, 10, 11, 12, or 13.
Claim 17
Dependent Claim
30 words
A microorganism transformed by a cloning vehicle of claims 7, 8, 9, 10, 11, 12, or 13 wherein expression of said DNA fragment produces bioluminescence upon addition of an aldehyde.
Claim 18
Dependent Claim
6 words
A transformed microorganism E. coli RR1/pTB7.
Claim 19
Dependent Claim
6 words
A transformed microorganism E. coli RR1/pTB7.
References Cited (Backward Citations)
No citation records indexed.
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| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
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| 3.5 Years (Year 4) | 1989-04-07 | 1989-10-07 | 1990-04-08 | $2000.00 | Window Closed / Lapsed |
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