Dissertations / Theses on the topic 'Expression à la surface de la levure'
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Pruvost, Tiphanie. "Ingénierie moléculaire de la réactivité croisée inter-espèces d’anticorps thérapeutiques par Yeast Surface Display." Electronic Thesis or Diss., université Paris-Saclay, 2023. http://www.theses.fr/2023UPASQ074.
Success of the monoclonal antibodies as therapeutic tools is partly due to their high specificity for their targets. Because of this high specificity antibodies developed against a human target often fail to recognize this target in animals used as models in preclinical trials. Thus, the goal of this study is to develop a protein engineering method aiming at conferring an antibody the ability to recognize a same antigen belonging to different species. To do so, two antibodies recognizing the human LAMP1 protein but not the murine and simian LAMP1 are used as models. In this project an exhaustive mutagenesis (DMS) monitored by Yeast Surface Display (YSD) and flow cytometry. The first part describes how these technics are combined in order to identify the epitopes of two antibodies on human LAMP1. The amino acids of epitopes are compared to those of murine and simian LAMP1 to explain the lack of recongnition of this two proteins. The second part focuses on the engineering strategy developped on the two antibodies. The expression of the DMS libraries in YSD allowed to select single mutations improving the cross-reactivity on LMAP1 without affecting the functionality of the antibodies. These mutations have been combined in a second library that has been screened for promissing variants. The affinities of these variants for differents human et simian LAMP1 orthologs has been measured to confirm the succes of the strategy. For each antibody many cross-reactive variants have been obtained
Sivelle, Coline. "Conception et production d’anticorps anti-TNFa non immunogènes pour le traitement des maladies inflammatoires." Thesis, Université Paris-Saclay (ComUE), 2019. http://www.theses.fr/2019SACLS603.
Efficacy of anti-TNFα antibodies is well known to be affected by their immunogenicity. Some patients developp anti-drug antibodies (ADA) which can elicit adverse effects and neutralize the therapeutic protein. Adalimumab (Humira®), which is the most used anti-TNFα, is reported to be immunogenic for more than 30% of patients in some diseases. T-cell epitopes that account for its immunogenicity are mostly carried by regions implied in the interaction with TNFα. In the present work, we propose to remove T-cell epitopes from the antibody sequence while maintaining its functionality.To undertake this issue, this PhD project uses Yeast Surface Display (YSD) to monitor the affinity of the biologic during the mutagenesis process. To do so, a comparison of different expression formats of antibody in YSD has been performed in order to define the format that will be used for the deimmunization method. Then, the strategy chosen to reduce immunogenicity is based on T-cell epitopes removal. First, it merges deep mutational scanning and in silico HLA II binding prediction to identify substitutions deleterious for HLA II/T-cell epitopes interaction while neutral for the function of the biologic. Secondly, these substitutions were combined to obtain libraries pre-enriched with functional sequences. Mutants with reduced immunogenic potential were then selected from these libraries. Several mutants of adalimumab with reduced immunogenicity potential according to HLA II binding prediction were identified and caracterized. All of them show an increased affinity for TNFα associated with an improvement of activity of at least two fold in comparison to adalimumab
Kudla, Bernard. "Construction d'un vecteur d'expression pour la levure Schizosaccharomyces pombe." Paris 11, 1987. http://www.theses.fr/1987PA112080.
Klonowska, Agnieszka. "Expression hétérologue de laccases du champignon Marasmius quercophilus chez la levure Saccharomyces cerevisiae." Aix-Marseille 3, 2000. http://www.theses.fr/2000AIX30085.
The basidiomycete Marasmius quercophilus colonizes the decaying litter of the evergreen oak (Quercus ilex L. ). This mushroom uses several enzymes to degrade lignin in order to have an access to cellulose. In vitro, it has a strong phenol oxidation activity related to the expression of several laccase isoforms among which Lac1 is predominant. Using molecular oxygen to oxidise numerous substrates, these copper enzymes are at the heart of many biotechnological projects. From controled culture conditions, we purified and partially characterized three laccase isoforms other than Lac1. Four members of the multigenic family which encodes laccases were cloned and sequenced. Among these genes, lac1, the structural gene encoding the major isoform, was formaly identified. The complementary DNA corresponding to these genes were expressed in the yeast Saccharomyces cerevisiae. Two of them, cDNA17 (lac1) and cDNA20, lead to the synthesis of secreted active laccases
Lacour, Thierry. "Expression du cytochrome p45011 bovin dans la levure : identification d'un nouveau transporteur d'electrons." Université Louis Pasteur (Strasbourg) (1971-2008), 1997. http://www.theses.fr/1997STR13101.
Boutabout, Mansour. "La Rétrotranscriptase de l'élément Ty1 de la levure Saccharomyces cerevisiae : Clonage, expression, fidélité." Université Louis Pasteur (Strasbourg) (1971-2008), 2002. http://www.theses.fr/2002STR13157.
Kribii, Rachida. "La squalène synthase d'Arabidopsis thaliana - caractérisation moléculaire et expression chez la levure Saccharomyces cerevisiae." Poitiers, 1996. http://www.theses.fr/1996POIT2390.
CORREDOR, RODRIGUEZ MAURICIO. "Diversite genetique, constitution chromosomique et expression genetique chez la levure fromagere halotolerant debaryomyces hansenii." Paris 11, 2001. http://www.theses.fr/2001PA112044.
Helies-Toussaint, Cécile. "Expression des genes dans le testicule." Paris 11, 1996. http://www.theses.fr/1996PA11T012.
Verdiere, Jacqueline. "Régulation de la synthèse de l'iso 2-cytochrome c chez la levure Saccharomyces cerevisiae." Paris 11, 1987. http://www.theses.fr/1987PA112281.
Daudonnet, Sylvie. "Transfert et expression du gène de la farnésyl diphosphate synthase de levure chez le tabac." Poitiers, 1996. http://www.theses.fr/1996POIT2264.
FLEURY, CHRISTOPHE. "Etude fonctionnelle des proteines decouplantes mitochondriales (ucps) par expression heterologue dans la levure saccharomyces cerevisiae." Paris 11, 1998. http://www.theses.fr/1998PA112237.
Worapamorn, Wilairat. "Cell-surface proteoglycan expression by periodontal cells /." St. Lucia, Qld, 2001. http://www.library.uq.edu.au/pdfserve.php?image=thesisabs/absthe16097.pdf.
Cool, Marc. "Etude des gènes codant pour le facteur d'élongation EF-1 alpha de levure : isolement et analyse du gène TEF2, contribution à la construction de vecteur d'expression de gène hétérologue à la levure." Paris 6, 1986. http://www.theses.fr/1986PA066363.
Morris, L. "Expression of surface molecules on mouse foetal macrophages." Thesis, University of Oxford, 1988. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.235069.
Lutton, Deborah A. "Variability of surface structure expression in Bacteroides fragilis." Thesis, Queen's University Belfast, 1991. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.334571.
Al-Subaie, Abeer. "Genetic studies on surface expression of platelet glycoproteins." Thesis, University of Cambridge, 2014. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.648804.
Bretes, Rodrigues Hugo. "Etude de la régulation du métabolisme des ARN messagers chez la levure Saccharomyces cerevisiae." Thesis, Paris 11, 2012. http://www.theses.fr/2012PA112209.
During transcription, several factors associate with mRNA to form messenger Ribonucleoparticles (mRNPs), thereby controlling their processing, their stability, and their cytoplasmic fate. To ensure the production of functional proteins from these mRNAs, eukaryotic cells contain numerous regulatory and quality control systems in order to prevent aberrant mRNP accumulation and export.In the yeast Saccharomyces cerevisiae, several nuclear pore associated proteins, including the SUMO isopeptidase Ulp1, have been involved in a mRNP quality control regulating their nuclear export. These data suggested that post-translational modification by SUMO of one or several mRNP components could regulate mRNA export. In order to understand the molecular mechanisms underlying this process, we undertook several approaches to identify these SUMOylated factors. In particular, we have set up a proteomic screen to identify mRNP components whose assembly onto mRNPs depends on Ulp1 activity.This proteomic survey revealed an Ulp1-dependent regulation of THO complex assembly to mRNPs. This complex, recruited to transcribed genes and mRNPs, is known to regulate transcription elongation by preventing DNA-RNA hybrids formation (termed R-loops), and mRNP export. Through a combination of proteomic analysis of mRNPs assembled in Ulp1 mutant cells, with RNA / chromatin immunoprecipitation experiments, we demonstrate that Ulp1 controls specifically the recruitment of the THO complex within mRNPs. SUMOylation analysis further reveals that Ulp1 targets the THO complex subunit Hpr1 on its C-terminal domain for deSUMOylation. We further show that this SUMOylation event regulates THO complex association within mRNPs. Finally, functional analysis reveal that impaired deSUMOylation of the THO complex do not affect mRNP export, but disturbs expression of LacZ reporter genes, a phenotype classically associated with THO complex dysfunction. Intriguingly, the transcriptional effect of inactivation or impaired deSUMOylation of the THO complex on LacZ expression is alleviated by the presence of an intron, providing a molecular basis for previously reported pre-mRNA leakage phenotypes. Our data therefore unravels for the first time a function of SUMO in the control of mRNP assembly contributing to proper mRNP homeostasis
Sarramegna, Valérie. "Solubilisation et purification du récepteur u-opoi͏̈de humain surexprimé dans la levure méthylotrophe pichia pastoris." Toulouse 3, 2002. http://www.theses.fr/2002TOU30034.
Enjalbert, Brice. "Régulation transcriptionnelle de Gsy2p, glycogène synthase majeure de la levure Saccharomyces cerevisiae." Toulouse, INSA, 2001. http://www.theses.fr/2001ISAT0003.
The Yeast Saccharomyces cerevisiae, like most of the living organisms, has to cope with stress and environmental constraints to ensure its survival and proliferation. One of this adaptive response is the synthesis of glycogen during a nutritional limitation. We have demonstrated that the whole group of genes allowing the glycogen synthesis or degradation is induced at the same time, before the end of the exponential phase of growth on glucose. Most of these genes possess STREs (STress Response Elements) in their promoter which are responsible for their increase in expression due to a stress but not to the disappearance of glucose. This induction requires at least two elements in the promoter of the GSY2 gene, encoding the major glycogen synthase. Study of the main mutations affecting GSY2 expression has led to the unraveling of a fine regulation system implicating three nutritional signaling pathways. The elements regulating the GSY2 transcription have been localized and the cAMP/PKA has been demonstrated to be the main pathway controlling the gene expression and induction. Moreover, our work on the linkage between glycogen biosynthesis and respiratory requirement have allowed to precise the production and consumption kinetics of glycogen and its function as a sugar storage
PEYRONNEAU, MARIE-ANNE. "Expression et caracterisation biochimique et spectroscopique de p450s humains de la famille 3a dans la levure saccharomyces cerevisiae." Paris 7, 1994. http://www.theses.fr/1994PA077076.
Lemieux, Caroline. "Caractérisation de l'homologue de PABPN1 (Poly(A)-Binding Protein Nuclear 1) chez la levure à fission Schizosaccharomyces pombe." Thèse, Université de Sherbrooke, 2012. http://hdl.handle.net/11143/6241.
Tavares, Rafaela. "Surface expression of Chromobacterium violaceum transaminase in Escherichia coli." Thesis, KTH, Skolan för bioteknologi (BIO), 2011. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-49131.
Brousse, Michel. "Analyse qualitative et quantitative du potentiel fermentaire de quelques souches industrielles de Levure et de son expression au cours de la fermentation alcoolique du moût de raisin." Bordeaux 2, 1985. http://www.theses.fr/1985BOR22013.
LOULERGUE, CLARISSE. "Caracterisation d'adnc de racines de mais par expression dans des mutants de levure alteres dans le transport du fer." Paris 7, 1998. http://www.theses.fr/1998PA077252.
Hocquet, Clémence. "Etude du rôle de Condensine dans le contrôle de l'expression génique chez la levure Schizosaccharomyces pombe." Thesis, Lyon, 2018. http://www.theses.fr/2018LYSEN037.
Condensin is a genome organiser that shape chromosomes and promote their accurate transmission in anaphase. Several studies have related changes in RNA level when Condensin is defective, suggesting that the complex has also a role in gene expression. However, the mechanisms have remained enigmatic and we still don’t know to what extent it is related to its role in chromosome organization. During my thesis, I studied the role played by Condensin in the regulation of gene expression using S. pombe as a model system. In contrast to previous studies, my results provide compelling evidence that Condensin plays no direct role in the maintenance of the transcriptome, neither during interphase nor during mitosis in this yeast. Accordingly to previous studies, I observed changes in RNA level in cells mutated for Condensin; non coding and 3’ extended RNA accumulate. However, I showed that the changes in gene expression in post-mitotic fission yeast cells that result from Condensin inactivation are largely a consequence of chromosome missegregation during anaphase, which notably depletes the RNA-exosome from daughter cells. Crucially, preventing karyotype abnormalities in daughter cells restores a normal transcriptome despite Condensin inactivation. Thus, chromosome instability, rather than a direct role of Condensin in the transcription process, changes gene expression. This work challenges the concept of gene regulation by canonical Condensin complexes and ask for caution when studying Condensin role outside chromosome condensation in mitosis
Wong, Angela Ting Ting. "The effect of surface topography on gene expression of macrophages." Thesis, University of British Columbia, 2013. http://hdl.handle.net/2429/45058.
Cresswell, Joanne. "HIV modulation of MHC class II biosynthesis and surface expression." Thesis, University of Edinburgh, 1999. http://hdl.handle.net/1842/14742.
Boron, Mallorie Lynn. "Expression and Cell Surface Re-Engineering of Thrombomodulin on Macrophages." Cleveland State University / OhioLINK, 2020. http://rave.ohiolink.edu/etdc/view?acc_num=csu1588167014316672.
Storlie, Patricia Ann. "Expression of the major surface protease (MSP) of leishmania chagasi." Diss., University of Iowa, 2009. https://ir.uiowa.edu/etd/891.
Lagorce, Arnaud. "Etude du mécanisme de compensation induit en réponse à des mutations pariétales chez la levure Saccharomyces cerevisiae." Toulouse, INSA, 2002. http://www.theses.fr/2002ISAT0021.
Saccharomyces cerevisiae cells are surrounded by a cell wall, which consists of a complex structure essentially composed of polymers of glucose units (b-glucans), of mannose units (mannans) and of N-acetylglucosamine units (chitin). In order to investigate on the cell wall rescue mechanism induced in response to cell wall mutations, we studied five mutants affected in different pathways of the cell wall metabolism. First, this work demonstrated the key role played by the gene GFA1, which encodes a glutamine fructose-6P-amidotransferase, in the activation of the chitin biosynthesis pathway in response to cell wall mutation. Secondly, we investigated the cell wall rescue mechanism using high density filters arrays. This approach led to the characterisation of the cell wall compensatory transcriptional signature and to the identification of a novel regulatory motif named WCE (for Wall Consensus Element). Moreover, promoter analysis of the genes implicated in the cell wall compensatory mechanism clarified the two regulatory pathways underlying this mechanism : the cell wall integrity pathway and the general stress response pathway
PIGNEDE, GEORGES, and Giuseppe Baldacci. "Structure, fonction et expression du gene de la sous-unite catalytique de l'adn polymerase delta de la levure schizosaccharomyces pompbe." Paris 6, 1993. http://www.theses.fr/1993PA066205.
Li, Tong-Tong. "Molecular biology of cell reactions to surface topography." Thesis, University of Glasgow, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.312506.
Montigny, Jacky de. "URA5 et URA10, deux gènes codant pour deux isoenzymes à activité OMP pyrophosphorylase chez la levure Saccharomyces cervisiae structure, expression, régulation /." Grenoble 2 : ANRT, 1988. http://catalogue.bnf.fr/ark:/12148/cb376166778.
Sheader, Karen. "The architecture and control of variant surface glycoprotein gene expression sites." Thesis, University of Oxford, 2004. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.403985.
Miller, Erin Suzanne. "Increasing Expression of Hepatitis B Surface Antigen in Maize through Breeding." DigitalCommons@CalPoly, 2015. https://digitalcommons.calpoly.edu/theses/1359.
Prudent, Sébastien. "Expression de l'aquaporine tonoplastique Bob TIP1 ;1 de chou-fleur chez la levure Saccharomyces cerevisiae : rôle dans la réponse cellulaire au stress hypo-osmotique." Dijon, 2005. http://www.theses.fr/2005DIJOS026.
Osmoregulation plays an important role in cellular responses to osmotic stress in plants and in yeast. Aquaporins contribute to osmotic adjustment by facilitating transport of water or solutes across membranes. The tonoplastic water channel BobTIP1;1 genes are upregulated during dessication stress in cauliflower meristematic tissue. To investigate the physiological importance of BobTIP1;1, we expressed it in a Saccharomyces cerevisiae osmosensitive mutant fps1. We showed that the defect in the yeast glycerol plasma membrane transporter transporter is complemented by a plant cDNA encoding the aquaporin BobTIP1;1 which is localized in the vacuolar membrane of the complemented yeast cells. To our knowledge, this is the first example of a plant aquaporin for which localization in the vacuolar membrane of yeast cells is related to an osmoresistant phenotype under hypo-osmotic shock
Fégueur, Marc. "Régulation de l'activité squalène synthétase chez la levure saccharomyces cerevisiae : isolement du gene et détermination des modalités de régulation de son expression." Poitiers, 1991. http://www.theses.fr/1991POIT2317.
Bordonné, Rémy. "Structure et expression de genes mitochondriaux de la levure saccharomyces cerevisiae : etude de la maturation des produits de transcription du dna mitochondrial." Université Louis Pasteur (Strasbourg) (1971-2008), 1987. http://www.theses.fr/1987STR13187.
Montigny, Jacky de. "Ura5 et ura10, deux genes codant pour deux isoenzymes a activite omp pyrophosphorylase chez la levure saccharomyces cerevisiae : structure, expression et regulation." Strasbourg 1, 1988. http://www.theses.fr/1988STR13198.
Bordonné, Rémy. "Structure et expression de gènes mitochondriaux de la levure Saccharomyces cerevisiae étude de la maturation des produits de transcription du DNA mitochondrial /." Grenoble 2 : ANRT, 1987. http://catalogue.bnf.fr/ark:/12148/cb376031669.
Li, Zhongyan. "Characteristics of platelet surface expression of glycoprotein VI in type 2 diabetes." [S.l.] : [s.n.], 2004. http://deposit.ddb.de/cgi-bin/dokserv?idn=972317430.
Chou, Laisheng. "Effects of substratum surface chemistry and topography on extracellular matrix gene expression." Thesis, National Library of Canada = Bibliothèque nationale du Canada, 1997. http://www.collectionscanada.ca/obj/s4/f2/dsk3/ftp04/nq25033.pdf.
Nazzari, Hamed. "Structural determinants regulating surface expression and function of Kv-related ion channels." Thesis, University of British Columbia, 2010. http://hdl.handle.net/2429/28410.
So, Ling-yue Daphne, and 蘇令如. "Inclined [expression], [impression]: an urbanconnector/collector on the inclined surface at foothill." Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 2001. http://hub.hku.hk/bib/B31986031.
Jarmander, Johan. "Improved detection and performance of surface expression from the AIDA-I autotransporter." Licentiate thesis, KTH, Bioprocessteknik (stängd 20130101), 2013. http://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-121561.
QC 20130506
Crummie, Darragh Kevin. "Modulation of NMDA receptor surface expression by DISC1 and its pathway partners." Thesis, University of Edinburgh, 2015. http://hdl.handle.net/1842/21047.
Trimnell, Adama R. "Assessment of cell surface expression vectors for heterologous expression of peptides/antigens through the A-layer of Aeromonas salmonicida." Thesis, University of Reading, 1996. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.339514.
Vincenti, Sophie. "Expression de l’hydroperoxyde lyase recombinante d’olive chez la bactérie Escherichia coli et chez la levure Pichia pastoris Etude des relations structure-fonction de l’enzyme." Thesis, Corte, 2016. http://www.theses.fr/2016CORT0010.
Hydroperoxide lyase (HPL) is an enzyme found in higher plants and involved in the lipoxygenase pathway. HPL acts on the 9- and/or 13-hydroperoxy fatty acids to produce C6 or C9 aldehydes, responsible for the fresh odor of cut grass known as "green note" and widely used by cosmetic and food industries. Production systems using recombinant enzymes are developed to meet the growing demand for natural green leaf volatiles. Thus, HPL from olive fruit (HPLwt) and the enzyme with its chloroplast transit peptide deleted (HPLdel), were cloned in the laboratory and expressed in Escherichia coli. Both recombinant enzymes were purified and biochemically characterized. They act only on 13-hydroperoxides, exhibit maximum activity at pH 7.5 and 25°C, and are more effective on the 13-hydroperoxy linoleic acid. The study of HPLwt and HPLdel stability over time has shown that all of the enzymatic activity is retained during a five weeks storage at -80°C in the presence of 10% glycerol. Furthermore, the addition of chemical compounds such as NaCl, Na2SO4 and glycine have increased the activity of both enzymes. Meanwhile, recombinant HPLs have been expressed in the eukaryotic system Pichia pastoris yeast. The HPLs are not secreted into the culture medium, but are expressed intracellularly, with a maximum after 24h of induction. Moreover, HPL structure-function relations were studied. The olive 13-HPL three dimensional structure has been built by computational modelling. Amino acids potentially involved in regiospecificity of the enzyme have been identified and modified by site-directed mutagenesis. Thus, T302K mutant has a catalytic efficiency on 13-hydroperoxides similar to the HPLwt, but it is also active on the 9-hydroperoxide of linoleic acid
Bednarska, Aleksandra. "Artificial systems for in vitro gene expression." Thesis, Université Paris-Saclay (ComUE), 2015. http://www.theses.fr/2015SACLN016/document.
DNA-dependent RNA polymerase (RNAP) is an enzyme responsible for the polymerization of ribonucleotides into an RNA sequence complementary to the template DNA. RNAP family has several members being single subunit (e.g. T7 bacteriophage) or multi subunit (bacterial and eukaryote) proteins. RNA transcription – a crucial event in gene expression – differs depending on the RNAP origin. Although the transcription process is relatively well characterized, many elements remain poorly understood, especially with respect to the dynamics of promoter recognition, escape and elongation in a cell like context where molecular density, concentrations and nearest neighbour effects are prevalent.The goal of this thesis was to develop a robust method that would allow real time monitoring of RNAP reaction in vitro in thoroughly controlled conditions. A major axis was to develop a surface-based biosensor that would allow the characterization of the main steps of the transcription reaction. Consequently, interactions between DNA molecules immobilized on a sensor surface and free RNAP delivered through a microfluidic flow system to the surface were examined. Changes in refractive index, correlated with changes in mass at a surface were followed using surface plasmon resonance imaging (SPRi). SPRi is a sensitive technique dedicated to analysis of interactions between two ligands in real time. The mechanism bases on the detection of slight differences in the reflectivity of polarized light at a fixed angle that are associated with a mass variation at the interface. Data obtained from SPRi are used to determine the kinetics of the interactions. Microarray geometry of SPRi allows monitoring several samples simultaneously that significantly shortens manipulation time and improves a quality and reproducibility of obtained results. Other label-free optofluidic biosensors: microring resonator and total internal reflection fluorescence (TIRF) microscopy were developed in parallel.We firstly biofunctionalized and characterized sensor surfaces (polymer coated glass for microring resonator and TIRF microscopy and 50-nm thin layer gold coatings on glass prisms for SPRi) in order to immobilize DNA strands in a controlled manner, using a self-assembled monolayer (SAM). Functionalization of photoresist polymer SU-8 concerned two methods: covalent (bio)molecule grafting and non-covalent conjugation based on hydrophobic coupling. Regarding gold surface functionalization, four different strategies of antifouling (bio)molecule immobilization were compared: thiol – gold bond formation, amide bond formation, extrAvidin – biotin interactions and hydrophobic coupling. Studies of DNA conjugation to the functionalized gold surface were performed with respect to specificity and density of immobilized DNA molecules of different lengths: 50, 500 and 1000 bp.Finally, biofunctionalized surfaces were used for real time monitoring of transcription reactions using two RNAPs: monomeric bacteriophage T7 RNAP and the holoenzyme of Escherichia coli RNAP. Kinetic analyses of nucleoprotein complex formation and RNA transcription were performed as a function of immobilized DNA density, the length of the immobilized DNA, the position of the specific promoter sequence with respect to the point of immobilization and the direction of subsequent transcription. RNA transcription in the SPRi apparatus was confirmed by collection, detection and analysis of relevant products.The future development of biosensors dedicated to in vitro gene expression will include the adaptation of the methods presented above to other optofluidic systems and further development of the technique. The final goal comprises a controlled RNA synthesis that would be an intermediate step to investigate real time in vitro protein production