subject:(renilla luciferase)

University of Helsinki

1. Titov, Soubir. Protein-protein interaction assay with split luciferase in planta.

Degree: Department of Agricultural Sciences; Helsingfors universitet, Agrikultur- och forstvetenskapliga fakulteten, Institutionen för lantsbruksvetenskaper, 2010, University of Helsinki

URL: http://hdl.handle.net/10138/17553

► Protein-protein interactions (PPIs) regulate many different cellular processes including transcription, translation, cell division, signal transduction, and oncogenic transformation. It is therefore important to develop sensitive… (more)

▼ Protein-protein interactions (PPIs) regulate many different cellular processes including transcription, translation, cell division, signal transduction, and oncogenic transformation. It is therefore important to develop sensitive and versatile techniques for the detection of these protein-protein interactions in order to fully understand protein functions. The most commonly used and traditional technique, the yeast two-/three hybrid (Y2H/Y3H) method, often results in false positives and false negatives, and other widely used techniques, such as bioluminescence resonance energy transfer (BRET), fluorescence resonance energy transfer (FRET), and bimolecular fluorescence complementation (BiFC) require extensive instrumentation. When compared with other PPI detection methods, the luciferase-based complementation assay specially split luciferase is believed to deliver the most sensitive and highest dynamic range, making it ideal for large-scale analysis. Therefore, for testing PPIs in planta, split Renilla luciferase complementation assay was chosen. In order to conduct this experiment, a series of plasmid constructs were made to enable the transient expression of fusion proteins. A well known protein pair, Arabidopsis nuclear Histone 2A and 2B, was tested initially as a proof of concept, and then three more proteins of the Gerbera MADS-box B class were investigated. For Arabidopsis Histone 2A and 2B, the intensity in all combinations was on average 9.4-fold higher in Relative Luminescence Units (RLUs) than the mock treated protoplasts. Moreover, in the case of Gerbera MADS-box B class proteins, the protein pairs GDEF1-GDEF2, GDEF1-GGLO1, and GDEF2-GGLO1 showed 8.4-19.4, 9.5-15.8, and 8.3-9.1-fold higher signals than the mock treated protoplasts. These results suggest that various complexes formed from different combinations of these three B class MADS-box proteins may increase the complexity of their regulatory functions, thus specifying the molecular basis of whorl morphogenesis and combinatorial interactions of floral organ identity genes in Gerbera. Finally, it was concluded that split Renilla luciferase can be a simple, reliable, fast, and effective method for examining PPIs in planta.

Subjects/Keywords: protein-protein interaction; split Renilla luciferase; MADS-box genes; flower development; vector construction; polyethylene glycol; electroporation; luminometer; Bioteknik (MAAT); Biotechnology (MAAT); Biotekniikka (MAAT); protein-protein interaction; split Renilla luciferase; MADS-box genes; flower development; vector construction; polyethylene glycol; electroporation; luminometer

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Titov, S. (2010). Protein-protein interaction assay with split luciferase in planta. (Masters Thesis). University of Helsinki. Retrieved from http://hdl.handle.net/10138/17553

Chicago Manual of Style (16^th Edition):

Titov, Soubir. “Protein-protein interaction assay with split luciferase in planta.” 2010. Masters Thesis, University of Helsinki. Accessed December 14, 2019. http://hdl.handle.net/10138/17553.

MLA Handbook (7^th Edition):

Titov, Soubir. “Protein-protein interaction assay with split luciferase in planta.” 2010. Web. 14 Dec 2019.

Vancouver:

Titov S. Protein-protein interaction assay with split luciferase in planta. [Internet] [Masters thesis]. University of Helsinki; 2010. [cited 2019 Dec 14]. Available from: http://hdl.handle.net/10138/17553.

Council of Science Editors:

Titov S. Protein-protein interaction assay with split luciferase in planta. [Masters Thesis]. University of Helsinki; 2010. Available from: http://hdl.handle.net/10138/17553

Université Paris-Sud – Paris XI

2. Gealageas, Ronan. Synthèse d’analogues de la coelentérazine pour l’imagerie in vivo dynamique des signaux calciques - Synthèse d’analogues du (-)-EGCG comme inhibiteurs de Dyrk1a dans la thérapie symptomatique de la trisomie 21 : Synthesis of coelenterazine analogs for dynamic in vivo imaging of calcium signaling -Synthesis of (-)-EGCG analogs as Dyrk1a inhibitors in the symptomatic therapy of Down syndrome.

Degree: Docteur es, Chimie, 2011, Université Paris-Sud – Paris XI

URL: http://www.theses.fr/2011PA112264

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Au cours de cette thèse, deux sujets distincts ont été étudiés : d’une part la synthèse d’analogues de la coelentérazine, substrat de différentes luciférases, pour… (more)

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Au cours de cette thèse, deux sujets distincts ont été étudiés : d’une part la synthèse d’analogues de la coelentérazine, substrat de différentes luciférases, pour une application en imagerie in vivo, et d’autre part la synthèse d’analogues du (-)-EGCG, inhibiteur de la kinase Dyrk1a, impliquée notamment dans les troubles cognitifs rencontrés dans la trisomie 21.La problématique du premier sujet consistait à obtenir des analogues de la coelentérazine conservant leur activité sur deux luciférases, la luciférase Renilla et l’aequorine, tout en induisant un déplacement vers le rouge de la bioluminescence produite par ces enzymes. L’aequorine, sensible au calcium, représentait la cible biologique principale du projet.Sept analogues dont six originaux ont été obtenues par des méthodes de synthèse classiques, et leurs activités ont pu être testées sur les deux luciférases choisies, avec des résultats probants : malgré des émissions de lumière moins intenses que celles obtenues avec la coelentérazine native, plusieurs molécules ont entrainé un déplacement vers le rouge de la longueur d’onde d’émission de lumière par bioluminescence allant jusqu’à 27 nm pour l’aequorine et plus de 120 nm pour la luciférase Renilla.Le second sujet, de chimie médicinale classique, a principalement consisté à la synthèse d’analogues du gallate d’épigallocatéchine (EGCG) au squelette simplifié, et au sein desquels le cycle pyranique caractéristique des catéchines a été remplacé par un carbocycle. Plusieurs molécules ont pu être synthétisées, dont deux présentant le motif hexaphénol. Leur activité inhibitrice de Dyrk1a a pu être testée in vitro et l’une d’entre elle s’est déjà révélée plus active que l’EGCG.

During this thesis, two distinct projects were studied: on the one hand, the synthesis of coelenterazine analogs, substrate of several luciferases, in the purpose of using them for in vivo imaging, and on the other, the synthesis of (-)-EGCG analogs, inhibitor of the Dyrk1a kinase, which interests us for the role it plays in the mental retardation existing in the Down Syndrome disease.The problematic of the first project consisted in obtaining coelenterazine analogs that would not only maintain their activity on two luciferases, the Renilla luciferase and aequorine, but they should also induce a red-shift of the bioluminescence produced by these enzymes. Because of its sensitivity to calcium, aequorine was the main biologic target of this project.Seven analogs, of which six had an original structure, were synthesized through usual synthetic methodologies and their activities on both aequorine and Renilla luciferase were tested in vitro, with interesting results: even if the intensities of light emission were weaker than those obtained with native coelenterazine, several molecules produced a red-shift of the emission wavelength of bioluminescence, up to 27nm for aequorine and more than 120nm for the Renilla luciferase. The second project, of classical medicinal chemistry, mainly consisted in the synthesis of epigallocatechin gallate analogs…

Advisors/Committee Members: Dodd, Robert Hugh (thesis director).

Subjects/Keywords: Chimie médicinale; Coelentérazine; EGCG; Dyrk1a; Aequorine; Renilla; Luciférase; Bioluminescence; Chimioluminescence; Kinase; Suzuki; Déméthylation; Mitsunobu; Benzyne; Cétoacétal; Aminopyrazine; Medicinal chemistry; Coelenterazine; EGCG; Dyrk1a; Aequorine; Renilla; Luciferase; Bioluminescence; Chemiluminescence; Kinase; Suzuki; Demethylation; Mitsunobu; Benzyne; Ketoacetal; Aminopyrazine

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Gealageas, R. (2011). Synthèse d’analogues de la coelentérazine pour l’imagerie in vivo dynamique des signaux calciques - Synthèse d’analogues du (-)-EGCG comme inhibiteurs de Dyrk1a dans la thérapie symptomatique de la trisomie 21 : Synthesis of coelenterazine analogs for dynamic in vivo imaging of calcium signaling -Synthesis of (-)-EGCG analogs as Dyrk1a inhibitors in the symptomatic therapy of Down syndrome. (Doctoral Dissertation). Université Paris-Sud – Paris XI. Retrieved from http://www.theses.fr/2011PA112264

Chicago Manual of Style (16^th Edition):

Gealageas, Ronan. “Synthèse d’analogues de la coelentérazine pour l’imagerie in vivo dynamique des signaux calciques - Synthèse d’analogues du (-)-EGCG comme inhibiteurs de Dyrk1a dans la thérapie symptomatique de la trisomie 21 : Synthesis of coelenterazine analogs for dynamic in vivo imaging of calcium signaling -Synthesis of (-)-EGCG analogs as Dyrk1a inhibitors in the symptomatic therapy of Down syndrome.” 2011. Doctoral Dissertation, Université Paris-Sud – Paris XI. Accessed December 14, 2019. http://www.theses.fr/2011PA112264.

MLA Handbook (7^th Edition):

Gealageas, Ronan. “Synthèse d’analogues de la coelentérazine pour l’imagerie in vivo dynamique des signaux calciques - Synthèse d’analogues du (-)-EGCG comme inhibiteurs de Dyrk1a dans la thérapie symptomatique de la trisomie 21 : Synthesis of coelenterazine analogs for dynamic in vivo imaging of calcium signaling -Synthesis of (-)-EGCG analogs as Dyrk1a inhibitors in the symptomatic therapy of Down syndrome.” 2011. Web. 14 Dec 2019.

Vancouver:

Gealageas R. Synthèse d’analogues de la coelentérazine pour l’imagerie in vivo dynamique des signaux calciques - Synthèse d’analogues du (-)-EGCG comme inhibiteurs de Dyrk1a dans la thérapie symptomatique de la trisomie 21 : Synthesis of coelenterazine analogs for dynamic in vivo imaging of calcium signaling -Synthesis of (-)-EGCG analogs as Dyrk1a inhibitors in the symptomatic therapy of Down syndrome. [Internet] [Doctoral dissertation]. Université Paris-Sud – Paris XI; 2011. [cited 2019 Dec 14]. Available from: http://www.theses.fr/2011PA112264.

Council of Science Editors:

Gealageas R. Synthèse d’analogues de la coelentérazine pour l’imagerie in vivo dynamique des signaux calciques - Synthèse d’analogues du (-)-EGCG comme inhibiteurs de Dyrk1a dans la thérapie symptomatique de la trisomie 21 : Synthesis of coelenterazine analogs for dynamic in vivo imaging of calcium signaling -Synthesis of (-)-EGCG analogs as Dyrk1a inhibitors in the symptomatic therapy of Down syndrome. [Doctoral Dissertation]. Université Paris-Sud – Paris XI; 2011. Available from: http://www.theses.fr/2011PA112264

Universitat Pompeu Fabra

3. Guidi, Mònica. Micro RNA-Mediated regulation of the full-length and truncated isoforms of human neurotrophic tyrosine kinase receptor type 3 (NTRK 3).

Degree: Departament de Ciències Experimentals i de la Salut, 2009, Universitat Pompeu Fabra

URL: http://hdl.handle.net/10803/7114

► Las neurotrofinas y sus receptores constituyen una familia de factores cruciales para el desarrollo del sistema nervioso. La neurotrofina 3 ejerce su función principalmente a… (more)

Subjects/Keywords: RISC complex; retinoic acid; renilla luciferase; real-time quantitative PCR; PTBP1; pri-miRNA; pre-miRNA; post-transcriptional regulation; piRNA; PicTar; pGL4.13; PCR; P-body; p75NTR; overexpression; NTRK3; NTRK2; NTRK1; non-coding RNAs; NGF; neurotrophin; neurotrophic signaling; neuronal differentiation; neurodevelopment; neuroblastoma; nervous system; mRNA; miRNA target prediction; miRNA profiling; miRNA mimic; miRNA microarray; miRNA inhibitor; miRanda; microRNA; microarray; luciferase assay; Ingenuity Pathway Analysis (IPA); heLa cells; gene silencing; gene regulation; full-length isoform (FL-NTRK3); firefly luciferase; ERK; disease; dicer; cancer development; BDNF; argonaute; alternative splicing; 3'UTR; RNAhybrid; RT-PCR; SH-SY5Y cells; siRNA; standard error; synaptic plasticity; target validation; TargetScan; transfection; translational repression; TrkA; TrkB; TrkC; truncated isoform (TR-NTRK3); tyrosine kinase (TK); western blot; 575; 61

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Guidi, M. (2009). Micro RNA-Mediated regulation of the full-length and truncated isoforms of human neurotrophic tyrosine kinase receptor type 3 (NTRK 3). (Thesis). Universitat Pompeu Fabra. Retrieved from http://hdl.handle.net/10803/7114

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Guidi, Mònica. “Micro RNA-Mediated regulation of the full-length and truncated isoforms of human neurotrophic tyrosine kinase receptor type 3 (NTRK 3).” 2009. Thesis, Universitat Pompeu Fabra. Accessed December 14, 2019. http://hdl.handle.net/10803/7114.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Guidi, Mònica. “Micro RNA-Mediated regulation of the full-length and truncated isoforms of human neurotrophic tyrosine kinase receptor type 3 (NTRK 3).” 2009. Web. 14 Dec 2019.

Vancouver:

Guidi M. Micro RNA-Mediated regulation of the full-length and truncated isoforms of human neurotrophic tyrosine kinase receptor type 3 (NTRK 3). [Internet] [Thesis]. Universitat Pompeu Fabra; 2009. [cited 2019 Dec 14]. Available from: http://hdl.handle.net/10803/7114.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Guidi M. Micro RNA-Mediated regulation of the full-length and truncated isoforms of human neurotrophic tyrosine kinase receptor type 3 (NTRK 3). [Thesis]. Universitat Pompeu Fabra; 2009. Available from: http://hdl.handle.net/10803/7114

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

4. Πετράκης, Ευάγγελος. Νέες τεχνικές πρωτεϊνικής ανάλυσης.

Degree: 2007, University of Patras; Πανεπιστήμιο Πατρών

URL: http://hdl.handle.net/10442/hedi/17749

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In the first section of the present dissertation, a method is described for preparation of protein arrays. The method is based on the in vitro… (more)

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In the first section of the present dissertation, a method is described for preparation of protein arrays. The method is based on the in vitro expression of recombinant DNA sequences, which are produced via the polymerase chain reaction (PCR). Thus, a DNA array is converted into a protein array, facilitating the steps required for the production and isolation of proteins for immobilization. The second section presents a new nonradioactive approach to the protein truncation test. PCR is used as a synthetic tool for introduction of sequences that allow in vitro expression of the interrogated DNA and for its fusion in-frame upstream of a sequence encoding the photoprotein aequorin. In the absence of a mutation, active aequorin is generated after in vitro expression of the recombinant DNA molecule. On the contrary, any mutation that causes premature termination of translation results in no aequorin production. Finally, it is developed a rapid method for the preparation of biotinylated bioluminescent protein reporters. The method is based on the coupled in vitro transcription and translation of the corresponding DNA sequences, and the simultaneous biotinylation of the produced protein using a biotinylated analogue of the tRNA of lysine. The method was applied to the synthesis of biotinylated firefly luciferase and Renilla luciferase, avoiding the time-consuming steps of the interrogated cDNA’s expression in vivo, purification of the recombinant protein and conjugation with molecules which recognize the analyte.

Στο πρώτο μέρος της διδακτορικής διατριβής, αναπτύχθηκε μέθοδος παρασκευής συστοιχιών πρωτεϊνών, η οποία βασίζεται στην in vitro έκφραση μορίων DNA, που παράγονται με την αλυσιδωτή αντίδραση πολυμεράσης (PCR). Με τον τρόπο αυτό, μια συστοιχία DNA μετατρέπεται σε συστοιχία πρωτεϊνών, παρακάμπτοντας τα χρονοβόρα στάδια της κλωνοποίησης των επιθυμητών αλληλουχιών cDNA, της in vivo έκφρασής τους και της απομόνωσης των πρωτεϊνών. Στη δεύτερη ενότητα περιγράφεται μια νέα, μη ραδιοϊσοτοπική μέθοδος για τη δοκιμασία πρόωρου τερματισμού της πρωτεϊνοσύνθεσης. Στην προτεινόμενη μέθοδο, με τη χρήση της PCR ως συνθετικό εργαλείο, ενσωματώθηκε καθοδικά της εξεταζόμενης αλληλουχίας DNA η κωδικοποιούσα αλληλουχία της φωτοπρωτεΐνης ακουορίνης, διατηρώντας το ίδιο πλαίσιο ανάγνωσης. Μετά από έκφραση in vitro του ανασυνδυασμένου μορίου DNA, πραγματοποιείται άμεσος έλεγχος της ολοκλήρωσης ή μη της πρωτεϊνοσύνθεσης, μέσω της αντίδρασης της ακουορίνης, αποφεύγοντας τα πολύωρα στάδια της ηλεκτροφόρησης και της αυτοραδιογραφίας. Τέλος αναπτύχθηκε μέθοδος παρασκευής βιοτινυλιωμένων βιοφωταυγών ιχνηθετών, η οποία βασίζεται στην in vitro συζευγμένη μεταγραφή, μετάφραση και βιοτινυλίωση των παραγόμενων πρωτεϊνών. Έγινε σύνθεση των φωτοφερασών της πυγολαμπίδας και της Renilla, αποφεύγοντας τα στάδια κλωνοποίησης και έκφρασης των γονιδίων σε βακτήρια, της απομόνωσης των ανασυνδυασμένων πρωτεϊνών και της σύζευξής τους με μόρια αναγνώρισης του αναλύτη.

Subjects/Keywords: Συστοιχίες πρωτεϊνών; Μονονουκλεοτιδικοί πολυμορφισμοί; Δοκιμασία πρόωρου τερματισμού της πρωτεϊνοσύνθεσης; Βιοφωταυγείς πρωτεϊνες; Φωτοπρωτεϊνη ακουορίνη; Φωτοφεράση της πυγολαμπίδας; Φωτοφεράση της renilla; Συστήματα συζευγμένης μεταγραφής και μετάφρασης in vitro; Protein arrays; Single nucleotide polymorhisms; Protein truncation test; Bioluminescent proteins; Photoprotein aequorin; Firefly luciferase; In vitro transcription and translation systems

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APA · Chicago · MLA · Vancouver · CSE | Export to Zotero / EndNote / Reference Manager

APA (6^th Edition):

Πετράκης, . �. (2007). Νέες τεχνικές πρωτεϊνικής ανάλυσης. (Thesis). University of Patras; Πανεπιστήμιο Πατρών. Retrieved from http://hdl.handle.net/10442/hedi/17749

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Chicago Manual of Style (16^th Edition):

Πετράκης, Ευάγγελος. “Νέες τεχνικές πρωτεϊνικής ανάλυσης.” 2007. Thesis, University of Patras; Πανεπιστήμιο Πατρών. Accessed December 14, 2019. http://hdl.handle.net/10442/hedi/17749.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

MLA Handbook (7^th Edition):

Πετράκης, Ευάγγελος. “Νέες τεχνικές πρωτεϊνικής ανάλυσης.” 2007. Web. 14 Dec 2019.

Vancouver:

Πετράκης �. Νέες τεχνικές πρωτεϊνικής ανάλυσης. [Internet] [Thesis]. University of Patras; Πανεπιστήμιο Πατρών; 2007. [cited 2019 Dec 14]. Available from: http://hdl.handle.net/10442/hedi/17749.

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

Council of Science Editors:

Πετράκης �. Νέες τεχνικές πρωτεϊνικής ανάλυσης. [Thesis]. University of Patras; Πανεπιστήμιο Πατρών; 2007. Available from: http://hdl.handle.net/10442/hedi/17749

Note: this citation may be lacking information needed for this citation format:
Not specified: Masters Thesis or Doctoral Dissertation

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