CA2434737A1 - Non aggregating fluorescent proteins and methods for using the same - Google Patents
Non aggregating fluorescent proteins and methods for using the same Download PDFInfo
- Publication number
- CA2434737A1 CA2434737A1 CA002434737A CA2434737A CA2434737A1 CA 2434737 A1 CA2434737 A1 CA 2434737A1 CA 002434737 A CA002434737 A CA 002434737A CA 2434737 A CA2434737 A CA 2434737A CA 2434737 A1 CA2434737 A1 CA 2434737A1
- Authority
- CA
- Canada
- Prior art keywords
- nucleic acid
- protein
- chromo
- acid according
- aggregating
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 102000034287 fluorescent proteins Human genes 0.000 title claims abstract 8
- 108091006047 fluorescent proteins Proteins 0.000 title claims abstract 8
- 230000004931 aggregating effect Effects 0.000 title claims abstract 7
- 238000000034 method Methods 0.000 title claims 8
- 108020004707 nucleic acids Proteins 0.000 claims abstract 20
- 102000039446 nucleic acids Human genes 0.000 claims abstract 20
- 150000007523 nucleic acids Chemical class 0.000 claims abstract 20
- 108090000623 proteins and genes Proteins 0.000 claims abstract 12
- 102000004169 proteins and genes Human genes 0.000 claims abstract 12
- 239000012634 fragment Substances 0.000 claims abstract 3
- 230000009261 transgenic effect Effects 0.000 claims abstract 3
- 241000243321 Cnidaria Species 0.000 claims 5
- 108700019146 Transgenes Proteins 0.000 claims 2
- 230000002103 transcriptional effect Effects 0.000 claims 2
- 241000242757 Anthozoa Species 0.000 claims 1
- 108020004705 Codon Proteins 0.000 claims 1
- 125000000729 N-terminal amino-acid group Chemical group 0.000 claims 1
- 230000000977 initiatory effect Effects 0.000 claims 1
- 230000007935 neutral effect Effects 0.000 claims 1
- 239000002773 nucleotide Substances 0.000 claims 1
- 125000003729 nucleotide group Chemical group 0.000 claims 1
- 238000006467 substitution reaction Methods 0.000 claims 1
- 239000000203 mixture Substances 0.000 abstract 3
- 102000004196 processed proteins & peptides Human genes 0.000 abstract 2
- 108090000765 processed proteins & peptides Proteins 0.000 abstract 2
- 230000003993 interaction Effects 0.000 abstract 1
- 229920001184 polypeptide Polymers 0.000 abstract 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43595—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from coelenteratae, e.g. medusae
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Zoology (AREA)
- Biochemistry (AREA)
- Toxicology (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Gastroenterology & Hepatology (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Tropical Medicine & Parasitology (AREA)
- Peptides Or Proteins (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Nucleic acid compositions encoding non-aggregating chromo/fluoroproteins and mutants thereof, as well as the proteins encoded by the same, are provided. The proteins of interest are polypeptides that are non-aggregating colored and/or fluorescent proteins, where the the non-aggregating feature arises fr om the modulation of residues in the N-terminus of the protein and the chromo and/or fluorescent feature arises from the interaction of two or more residu es of the protein. Also provided are fragments of the subject nucleic acids and the peptides encoded thereby, as well as antibodies to the subject proteins and transgenic cells and organisms. The subject protein and nucleic acid compositions find use in a variety of different applications. Finally, kits for use in such applications, e.g., that include the subject nucleic acid compositions, are provided.
Claims (20)
1. ~A nucleic acid that encodes a non-aggregating chromo- or fluorescent mutant of an aggregating Cnidarian chromo- or fluorescent protein or mutant thereof.
2. ~The nucleic acid according to Claim 1, wherein said Cnidarian chromo-or fluorescent protein is from a non-bioluminescent Cnidarian species.
3. ~The nucleic acid according to Claim 2, wherein said non-bioluminescent Cnidarian species is an Anthozoan species.
4. ~A nucleic acid according to Claim 1, wherein said nucleic acid has a sequence of residues that is substantially the same as or identical to a nucleotide sequence of at least 10 residues in length of SEQ ID NOS:14; 15; 17; 19; 21;
and 23.
and 23.
5. ~A fragment of the nucleic acid selected according to Claim 1.
6. ~A construct comprising a vector and a nucleic acid according to Claim 1.
7. ~An expression cassette comprising:
(a) a transcriptional initiation region functional in an expression host;
(b) a nucleic acid according to Claim 1; and (c) and a transcriptional termination region functional in said expression host.
(a) a transcriptional initiation region functional in an expression host;
(b) a nucleic acid according to Claim 1; and (c) and a transcriptional termination region functional in said expression host.
8. ~A cell, or the progeny thereof, comprising an expression cassette according to Claim 7 as part of an extrachromosomal element or integrated into the genome of a host cell as a result of introduction of said expression cassette into said host cell.
9. ~A method of producing a chromo and/or fluorescent protein, said method comprising:
growing a cell according to Claim 8, whereby said protein is expressed; and isolating said protein substantially free of other proteins.
growing a cell according to Claim 8, whereby said protein is expressed; and isolating said protein substantially free of other proteins.
10. ~A protein or fragment thereof encoded by a nucleic acid according to Claim 1.
11. ~An antibody binding specifically to a protein according to Claim 10.
12. ~A transgenic cell or the progeny thereof comprising a transgene that is a nucleic acid according to Claim 1.
13. ~A transgenic organism comprising a transgene that is a nucleic acid according to Claim 1.
14. ~In an application that employs a chromo- or fluorescent protein, the improvement comprising:
employing a protein according to Claim 10.
employing a protein according to Claim 10.
15. ~In an application that employs a nucleic acid encoding a chromo- or fluorescent protein, the improvement comprising:
employing a nucleic acid according to Claim 1.
employing a nucleic acid according to Claim 1.
16. ~A kit comprising a nucleic acid according to Claim 1.
17. ~A method of producing a nucleic acid according to Claim 1, said method comprising:
modulating at least one N-terminal residue codon of an aggregating Cnidarian chromo and/or fluorescent protein encoding sequence to produce said nucleic acid.
modulating at least one N-terminal residue codon of an aggregating Cnidarian chromo and/or fluorescent protein encoding sequence to produce said nucleic acid.
18. ~The method according to Claim 17, wherein said at least one residue is a basic residue.
19. ~The method according to Claim 18, wherein said modulation is a substitution of said basic residue for a neutral residue.
20. ~The method according to Claim 18, wherein said basic residue is lys or arg.
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US27098301P | 2001-02-21 | 2001-02-21 | |
| US60/270,983 | 2001-02-21 | ||
| US10/006,922 | 2001-12-04 | ||
| US10/006,922 US7166444B2 (en) | 1999-10-14 | 2001-12-04 | Nucleic acids encoding chromophores/fluorophores and methods for using the same |
| PCT/US2002/005749 WO2002068459A2 (en) | 2001-02-21 | 2002-02-20 | Non aggregating fluorescent proteins and methods for using the same |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CA2434737A1 true CA2434737A1 (en) | 2002-09-06 |
| CA2434737C CA2434737C (en) | 2012-11-06 |
Family
ID=40863845
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA2434737A Expired - Lifetime CA2434737C (en) | 2001-02-21 | 2002-02-20 | Non aggregating fluorescent proteins and methods for using the same |
Country Status (5)
| Country | Link |
|---|---|
| EP (1) | EP1385967A2 (en) |
| JP (2) | JP2004536571A (en) |
| AU (1) | AU2002254031A1 (en) |
| CA (1) | CA2434737C (en) |
| WO (1) | WO2002068459A2 (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7005511B2 (en) * | 2001-02-26 | 2006-02-28 | The Regents Of The University Of California | Fluorescent protein variants and methods for making same |
| CA2439400A1 (en) * | 2001-02-26 | 2002-09-06 | The Regents Of The University Of California | Non-oligomerizing tandem fluorescent proteins |
| RU2330067C2 (en) | 2001-12-19 | 2008-07-27 | Дзе Юниверсити Оф Чикаго | Polynucleotide, coding chromo- or fluorescent mutant polypeptide, expression vector, cell, method of obtaining chromo- or fluorescent polypeptide, use of polypeptide and use of polynucleotide |
| RU2004118848A (en) * | 2001-12-26 | 2005-04-20 | Клонтек Лэборетериз, Инк. (Us) | MUTANT CHROMOPHORES / FLUOROPHORES AND METHODS FOR THEIR PRODUCTION AND APPLICATION |
| EP1567178A4 (en) * | 2002-11-01 | 2009-07-15 | Amgen Inc | Modulators of receptors for parathyrois hormone and parathyroid hormone-related protein |
| ATE443135T1 (en) | 2003-06-16 | 2009-10-15 | Riken | FLUORESCENT PROTEIN AND PIGMENT PROTEIN |
| EP1732944B1 (en) | 2004-04-07 | 2012-09-05 | The University of Chicago | Monomeric red fluorescent proteins |
| US8563703B2 (en) | 2006-01-25 | 2013-10-22 | Evrogen IP Joint Stock Company | Fluorescent proteins and methods for using same |
| RU2395581C2 (en) | 2006-01-25 | 2010-07-27 | Закрытое Акционерное Общество "Евроген Айпи" | Novel fluorescent proteins from entacmaea quadricolor and method of obtaining said proteins |
| JP2009533061A (en) * | 2006-04-14 | 2009-09-17 | セルセンス， インコーポレイテッド | Methods for assessing cell labeling |
| EP3575779A4 (en) * | 2017-01-27 | 2019-12-04 | Osaka University | METHOD FOR DETECTION OF BIOLOGICAL MATERIAL, AND CHEMILUMINESCENT INDICATOR USED IN THE METHOD |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU783767B2 (en) * | 1999-10-14 | 2005-12-01 | Takara Bio Usa, Inc. | Anthozoa derived chromophores/fluorophores and methods for using the same |
-
2002
- 2002-02-20 CA CA2434737A patent/CA2434737C/en not_active Expired - Lifetime
- 2002-02-20 EP EP02723238A patent/EP1385967A2/en not_active Withdrawn
- 2002-02-20 AU AU2002254031A patent/AU2002254031A1/en not_active Abandoned
- 2002-02-20 JP JP2002567969A patent/JP2004536571A/en not_active Withdrawn
- 2002-02-20 WO PCT/US2002/005749 patent/WO2002068459A2/en not_active Ceased
-
2008
- 2008-12-04 JP JP2008309300A patent/JP5221308B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JP5221308B2 (en) | 2013-06-26 |
| WO2002068459A2 (en) | 2002-09-06 |
| CA2434737C (en) | 2012-11-06 |
| JP2004536571A (en) | 2004-12-09 |
| JP2009131260A (en) | 2009-06-18 |
| EP1385967A2 (en) | 2004-02-04 |
| WO2002068459A3 (en) | 2003-11-27 |
| AU2002254031A1 (en) | 2002-09-12 |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| EEER | Examination request |