This paper closely examines Internet advertising techniques and tools which can be used to promote new product at the market. Goal of the thesis is to measure marketing campaign effectiveness and optimize advertising campaigns. Relevant KPIs were chosen to create evaluation matrices, identify successful advertising channels and create efficient internet marketing mix. Advertising activities described in the thesis helped Czech company to increase sales on the Internet. Practical part of the thesis describes process of introducing new product on the Czech market. In the initial stage several analyses were done to identify market conditions, competition and ideal consumer. Based on the results ¡V market entry strategy and web development plan were created. Internet e-shop and other supportive web pages were developed with aim to sell products through Internet channel. Websites follows best practice for web presentation design, search engine optimization and web audience measurement. During the project history several marketing campaigns were launched and the results were monitored using Google Analytics software. Selected marketing activities were closely examined. Concepts of Search engine optimization, POEM media, and Pay Per Click (PPC) advertising were used as field experiments. Partial results of each field experiment as well as overall results of all marketing activities are summarized in the conclusion. The thesis presents a comprehensive overview of the marketing tools and channels at the Czech market. Paper also summarizes best practices for website development and content optimization for visitors and search engines. Comparison of advertising activities in different channels within three PPC advertising networks helps to define PPC advertising strategy for Czech market. Based on the findings - optimized Internet marketing mix was created with aim to increase marketing campaigns effectiveness. Suggestions for optimization of website¡¦s content and recommendation for next marketing activities were summarized with aim to help company for future project development.

This study aims to shed light on the relationship between the commercial advertising model AIDA (Awareness/Attention, Interest, Desire, Action) and political television advertising, with a historical perspective being of extra interest. In order to do so, the study made use of theories concerning the AIDA-model, representation, rhetoric (with focus on ethos, pathos, and logos), and the professionalisation of political communication. The methodology involved qualitative analyses of 18 official political campaign advertisements from nine United States presidential elections between the years 1952-2016. One issue-ad from each candidate (Republicans and Democrats only) from every other election was strategically chosen for examination. Each advertisement was then analysed both as it relates to its rhetorical content as well as its structure with the defined four stages of the AIDA-model in mind, with any potential patterns between the rhetoric and the structure being taken into account. The results of the study suggest that while the AIDA-model can be recognised in political television advertisements in the United States since the inception in the 1950s, the advertisements from the post-modern phase of the professionalisation of political communication (1985-) seem to place more emphasis, compared to the modern phase (1950s-1985), on the desire stage of the AIDA-model. Furthermore, no distinct differences could be found between the parties from a pure rhetorical and structural standpoint, and both appear to be on practically identical evolutionary paths. An explanation to this could be the escalating reliance on hiring independent experts and specialist to manage the various areas involved with running a political campaign, which is a characteristic of the ever-increasingly professional environment of political communication.

Dagens digitala teknologi och sociala medier har alltmer satt sin prägel inom handeln och detta beror främst på att majoriteten människor idag har en smartphone där man ständigt är uppkopplad och aktiv inom sociala medier. Sociala medier används idag även av de flesta handelsföretag inom detaljhandeln och fungerar som en plattform där man kan nå ut till en större och bredare publik. Trenden växer och yttrar sig i att fler plattformar bildas och därav sätter press på småföretag för att kunna sticka ut bland mängden. I detta skede blir det därför allt viktigare för småföretag att vara aktiva på sociala medier för att väcka ett behov hos kunderna och fånga upp deras intresse vilket kan göras genom att publicera en variation av aktiviteter. Aktiviteterna kan inkludera marknadsföring med senaste nyheter och trender, tävlingar och även personliga inlägg. Detta engagemang ska fungera som en vägledning för att sedan locka och leda sina följare in till den fysiska butiken eftersom det är där köpet genomförs. Studien har utformats via en kvalitativ forskningsmetod med en abduktiv ansats som utgångspunkt. Studien innefattade totalt sex intervjuer varav fem av de var fysiskt på plats medan den sjätte var via telefon. Intervjuerna bidrog med det empiriska materialet i syfte att kunna få fram en analys där vi jämförde den teoretiska referensramen med det empiriska materialet. Ur analysen kunde vi se ett samband och resonerade fram vilka aspekter som sociala medier bidrog med för småföretag när de ska kommunicera sina aktiviteter och locka kunderna in till den fysiska butiken. Resultatet av studien yttrade sig i att sociala medier är ett viktigt hjälpmedel för småföretag då det inkluderar en gratis marknadsföring och att man kan nå ut till fler kunder och även utanför det geografiska området butiken befinner sig i. Kommunikation, interaktion och respons är viktiga faktorer för småföretag när de ska eller etablerar sig via sociala medieplattformar. Studiens resultat redogör även för småföretag vikten av att ständigt vara aktiv på sociala medier, publicera varierande aktiviteter, engagera sig i kunder genom att interagera och få feedback. När dessa aspekter interagerar ökar chansen för företagen att bygga långsiktiga kundrelationer. / Today's digital technology and social media have increasingly left their mark on commerce and this is mainly due to the fact that the majority of people today have a smartphone where they are constantly connected and active in social media. Today, social media is also used by most retail companies in the retail trade and acts as a platform where you can reach a larger and wider audience. The trend is growing and is reflected in the fact that more platforms are being formed and as a result put pressure on small companies to be able to stand out among the crowd. At this stage, it therefore becomes increasingly important for small companies to be active on social media in order to arouse a need in customers and capture their interest, which can be done by publishing a variety of activities. Activities can include marketing with the latest news and trends, contests and even personal posts. This commitment should serve as a guide to then attract and lead their followers into the physical store because that is where the purchase is made. The study has been designed via a qualitative research method with an abductive approach as a starting point. The study included a total of six interviews, five of which were physically in place while the sixth was by telephone. The interviews contributed with the empirical material in order to be able to obtain an analysis in which we compared the theoretical frame of reference with the empirical material. From the analysis, we could see a connection and reasoned out which aspects social media contributed to small companies when they have to communicate their activities and attract customers to the physical store. The result of the study was that social media is an important tool for small businesses as it includes free marketing and that you can reach more customers and also outside the geographical area the store is in. Communication, interaction and response are important factors for small businesses when they are going or establishing themselves via socialmedia platforms. The results of the study also explain to small companies the importance of being constantly active on social media, publishing various activities, engaging with customers by interacting and receiving feedback. When these aspects interact, companies' chances of building long-term customer relationships increase.

Syftet med denna studie är att undersöka hur digitala marknadsföringsbyråer arbetar medkontextuell annonsering och hur kontextuell annonsering påverkar kundens beslutsprocess.Teori om digitala marknadsföringsbyråer, kontextuell annonsering och kundens beslutsprocesshar tillämpats, samt en för undersökningen skapad analysmodell. Metoden som använts ärkvalitativ och har inkluderat fem digitalt utförda semi-strukturerade intervjuer.Undersökningen visar att digitala marknadsföringsbyråer arbetar med två former av kontextuellannonsering; tematisk annonsering och sökannonsering. Tematisk annonsering används främstför att uppnå varumärkeskännedom och sökannonsering används främst för att uppnå köpslut.Undersökningen visar vidare att tematisk annonsering påverkar kundens beslutsprocess genomatt väcka kundens uppmärksamhet och intresse för annonsen. Sökannonsering påverkarkundens beslutsprocess genom att väcka kundens önskan för produkten i annonsen och väckahandling genom att kunden köper en specifik produkt. Analysen förbättrar den nuvarandekunskapen om digitala marknadsföringsbyråers användning av kontextuell annonsering för attpåverka kundens beslutsprocess, och demonstrerar tillvägagångssätt som liknande aktörerskulle kunna anamma för att uppnå framgångsrik annonsering på internet.

Les autotransporteurs monomériques, appartenant au système de sécrétion de type V, correspondent à une famille importante de facteurs de virulence bactériens. Plusieurs fonctions, souvent essentielles pour le développement d’une infection ou pour le maintien et la survie des bactéries dans l’organisme hôte, ont été décrites pour cette famille de protéines. Malgré l’importance de ces protéines, notre connaissance de leur biogenèse et de leur mécanisme d’action demeure relativement limitée. L’autotransporteur AIDA-I, retrouvé chez diverses souches d’Escherichia coli, est un autotransporter multifonctionnel typique impliqué dans l’adhésion et l’invasion cellulaire ainsi que dans la formation de biofilm et d’agrégats bactériens. Les domaines extracellulaires d’autotransporteurs monomériques sont responsables de la fonctionnalité et possèdent pratiquement tous une structure caractéristique d’hélice β. Nous avons mené une étude de mutagenèse aléatoire avec AIDA-I afin de comprendre la base de la multifonctionnalité de cette protéine. Par cette approche, nous avons démontré que les domaines passagers de certains autotransporteurs possèdent une organisation modulaire, ce qui signifie qu’ils sont construits sous la forme de modules fonctionnels. Les domaines passagers d’autotransporteurs peuvent être clivés et relâchés dans le milieu extracellulaire. Toutefois, malgré la diversité des mécanismes de clivage existants, plusieurs protéines, telles qu’AIDA-I, sont clivées par un mécanisme qui demeure inconnu. En effectuant une renaturation in vitro d’AIDA-I, couplée avec une approche de mutagenèse dirigée, nous avons démontré que cette protéine se clive par un mécanisme autocatalytique qui implique deux acides aminés possédant un groupement carboxyle. Ces résultats ont permis la description d’un nouveau mécanisme de clivage pour la famille des autotransporteurs monomériques. Une des particularités d’AIDA-I est sa glycosylation par une heptosyltransférase spécifique nommée Aah. La glycosylation est un concept plutôt récent chez les bactéries et pour l’instant, très peu de protéines ont été décrites comme glycosylées chez E. coli. Nous avons démontré que Aah est le prototype pour une nouvelle famille de glycosyltransférases bactériennes retrouvées chez diverses espèces de protéobactéries. La glycosylation d’AIDA-I est une modification cytoplasmique et post-traductionnelle. De plus, Aah ne reconnaît pas une séquence primaire, mais plutôt un motif structural. Ces observations sont uniques chez les bactéries et permettent d’élargir nos connaissances sur la glycosylation chez les procaryotes. La glycosylation par Aah est essentielle pour la conformation d’AIDA-I et par conséquent pour sa capacité de permettre l’adhésion. Puisque plusieurs homologues d’Aah sont retrouvés à proximité d’autotransporteurs monomériques putatifs, cette famille de glycosyltranférases pourrait être importante, sinon essentielle, pour la biogenèse et/ou la fonction de nombreux autotransporteurs. En conclusion, les résultats présentés dans cette thèse apportent de nouvelles informations et permettent une meilleure compréhension de la biogenèse d’une des plus importantes familles de protéines sécrétées chez les bactéries Gram négatif. / Monomeric autotransporters, a family of proteins that use the type V secretion pathway, are important mediators of virulence for many bacterial pathogens. Many functions important for host colonization and survival have been described for these proteins. Despite the recognized importance of this family of proteins, the mechanisms that are required for the biogenesis and functionality of monomeric autotransporters still remain poorly understood. The Escherichia coli adhesin involved in diffuse adherence (AIDA-I) is a classical multifunctional autotransporter protein that mediates bacterial aggregation and biofilm formation, as well as adhesion and invasion of cultured epithelial cells. Extracellular domains of autotransporters are responsible for the protein function and fold into a characteristic β-helical structure. We performed a random mutagenesis of the AIDA-I passenger domain in order to identify regions involved in the various phenotypes associated with the expression of this protein. Our study suggests that the passenger domain of AIDA-I possesses a modular organization, which means that AIDA-I is built with individual functional modules. Autotransporter passenger domains can be cleaved from the β-domain and released into the extracellular milieu. However, despite the fact that diverse cleavage mechanisms have been previously described, many autotransporters, like AIDA-I, are cleaved by an unknown mechanism. By monitoring the in vitro refolding and cleavage following by site-directed mutagenesis, we showed that AIDA-I processing is an autocatalytic event that involves two acidic residues. Our results unveil a new mechanism of auto-processing in the autotransporter family. AIDA-I is one of the few glycosylated proteins found in Escherichia coli. Glycosylation is mediated by a specific heptosyltransferase encoded by the aah gene, but little is known about the role of this modification and the mechanism involved. Our findings suggest that Aah represents the prototype of a new large family of bacterial protein O-glycosyltransferases that modify various substrates recognized through a structural motif. Furthermore, we showed that glycosylation occurs in the cytoplasm by a cotranslational mechanism. These observations are unique in bacteria and represent a significant advance in our comprehension of prokaryotic glycosylation. We also showed that glycosylation is required to ensure a normal conformation of AIDA-I and, as a consequence, is necessary for its cell-binding function. The finding that other autotransporters or large adhesin-encoding genes are linked to Aah homologue-encoding genes suggests that glycosylation may be important, if not essential, for the function of these proteins, as for AIDA-I. In conclusion, the results presented in this thesis bring new information about the autotransporter family and also give new insight into the mechanisms that are important for different aspects of the biogenesis of monomeric autotransporters.

AIDA-I (adhesin involved in diffuse adherence) est une importante adhésine autotransporteur exprimée par certaines souches de Escherichia. coli impliquée dans la colonisation des porcelets sevrés causant la diarrhée post-sevrage et la maladie de l’œdème. Une précédente étude de notre laboratoire a identifié l’apolipoprotéine AI (ApoAI) du sérum porcin, la protéine structurale des lipoprotéines à haute densité, comme récepteur cellulaire putatif de AIDA-I. L’interaction entre ces deux protéines doit être caractérisée. Ici, nous montrons par ELISA que AIDA-I purifiée est capable d’interagir avec l’ApoAI humaine, mais également avec les apolipoprotéines B et E2. L’ApoAI est rencontrée sous deux formes, soit libre ou associée aux lipides. Nous montrons que la forme libre n’interagit pas avec les bactéries AIDA-I+ mais s’associe spécifiquement à l’ApoAI membranaire de cellules épithéliales HEp-2. Afin d’étudier le rôle de l’ApoAI dans l’adhésion des bactéries, nous avons infecté des cellules HEp-2 en présence d’anticorps dirigés contre l’ApoAI, mais l’adhésion des bactéries AIDA I+ n’a jamais été réduite. De plus, l’induction de l’expression de l’ApoAI par fénofibrate et GW7647 chez les cellules Caco 2 polarisée et Hep G2, n’a pas permis l’augmentation de l’adhésion cellulaire des E. coli exprimant AIDA-I. Notre étude suggère davantage que l’interaction entre AIDA-I et ApoAI n’intervient pas dans les mécanismes d’adhésion cellulaire. / The adhesin involved in diffuse adherence (AIDA-I) is an important autotransporter adhesin expressed by some strains of Escherichia coli and is involved in the intestinal colonisation of weaned piglets, causing the postweaning diarrhea and the edema disease. A previous study from our laboratory identified the apolipoprotein AI (ApoAI) from porcine serum, the structural protein of high density lipoproteins, as a putative receptor of AIDA-I. The interaction between these two proteins must be characterized. Here, we show that purified AIDA-I, using an ELISA assay, is able to bind the human ApoAI and the apolipoprotein B and E2. The ApoAI is found under two forms, either free or bound to lipid. We show that the free form of ApoAI does not interact with AIDA-I+ bacteria but specifically interact with membrane bound ApoAI on Hep-2 epithelial cells. To study the role of ApoAI in the adhesion of bacteria, we infected Hep-2 cells preincubated with antibodies to ApoAI. The adhesion of AIDA-I+ bacteria to the cells couldn’t be reduced. Additionally, the induction of ApoAI synthesis using fenofibrate and GW7647 on polarized Caco-2 or Hep G2 cells did not increase the adhesion of AIDA-I+ bacteria. Our study suggests that the interaction between AIDA-I and ApoAI is not involved in the cellular adhesion of the bacteria.

La question posée dans ce mémoire de recherche concerne l’artiste contemporain japonais Aida Makoto, comme figure provocante et ironique, remettant en question les appareils de répression et d’aliénation de la société capitaliste japonaise. L’objectif de ma réflexion est de montrer l’apport de la schizo-analyse dans l’analyse d’œuvres plastiques comportant des prédispositions à l’utilisation de celle-ci. À travers les œuvres de l’artiste Aida Makoto où une multiplicité de corps emplit l’espace de la toile, et à partir des concepts de multiplicité et de sensation théorisés par Gilles Deleuze et Félix Guattari, la recherche apportera une seconde lecture aux œuvres de cet artiste en mettant en avant les aspects révolutionnaires de sa création artistique. Constitué de deux chapitres, le mémoire porte dans un premier temps sur la picturalité de l’œuvre, d’ordre technique, esthétique et éthique, en mettant en avant les composés de sensation présents sur la toile, ceci afin, dans un second temps, d’appréhender la figuration, de la visagéité à la multiplicité, comme aspect central de l’œuvre. Ainsi, la Figure, au sens deleuzien du terme, permettrait à l’artiste Aida Makoto d’entamer une fuite schizophrénique à l’occasion de laquelle il pourra créer à l’abri de toute répression ou normalisation de ses machines désirantes par la société capitaliste japonaise. / The question posed in the research concerns the contemporary Japanese artist Aida Makoto, as a provocative and ironic figure, challenging the machinery of repression and alienation of the Japanese capitalist society. The purpose of my reflection is to show the contribution of schizoanalysis in the analysis of visual artworks. Through the artworks of the artist Aida Makoto, where a multiplicity of bodies fill the space of the canvas, and from the concepts of multiplicity and sensation, theorized by Gilles Deleuze and Félix Guattari, this research will bring a second reading to Aida’s works highlighting the revolutionary aspects of his artistic creation. Constituted of two chapters, the thesis focuses mainly on the pictoriality of the work from technical, aesthetic and ethics points of view, emphasizing the compounds of sensation present on the canvas, in order to, secondly, understand the figuration of faciality to multiplicity as a central aspect of the work. Thus the Figure, in the deleuzian sense, would allow the artist Makoto Aida to start a schizophrenic escape during which he can create freely from repression and normalization of desiring machines in the Japanese capitalist society.

This thesis deals with a comparatice analysis, where there are original literary productions (proses, dramas), that were then used for creating librettos of operas productions. The thesis focus on opera performances presented on stages of South Bohemian Theatre in last few years, and also belongs to canon of tradition of world's opera. The analysis is following the way of adaptation of the original piece, it means the conversion of the production, originally defined for a quiet reading, into the genre based on performing and accented his performance's nature. The thesis follows conventional and inventive characters by original literary production, and also by opera's text "adaptation". The thesis specifically follows the relationship of these productions: Victorien Sardou: Tosca and Giacommo Puccini: Tosca; Prosper Mérimée: Carmen and Georges Bizet: Carmen; Pierre - Augustin Caron de Beaumarchais: The Marriage of Figaro and Wolfgang Amadeus Mozart: The Marriage of Figaro; Antonio García Gutiérrez: Il Trovatore and Giuseppe Verdi: Il Trovatore; and also the productions without the original literar artwork: Giuseppe Verdi: Aida and Antonín Dvořák: Rusalka.

Escherichia coli (E. coli) is an attractive host for production of recombinant proteins, since it generally provides a rapid and economical means to achieve high product quantities. In this thesis, the impact of the glucose uptake rate on the production of recombinant proteins was studied, aiming at improving and optimising production of recombinant proteins in E. coli. E. coli can be cultivated to high cell densities in bioreactors by applying the fed-batch technique, which offers a means to control the glucose uptake rate. One objective of this study was to find a method for control of the glucose uptake rate in small-scale cultivation, such as microtitre plates and shake flasks. Strains with mutations in the phosphotransferase system (PTS) where used for this purpose. The mutants had lower uptake rates of glucose, resulting in lower growth rates and lower accumulation of acetic acid in comparison to the wild type. By using the mutants in batch cultivations, the formation of acetic acid to levels detrimental to cell growth could be avoided, and ten times higher cell density was reached. Thus, the use of the mutant strains represent a novel, simple alternative to fed-batch cultures.   The PTS mutants were applied for production of integral membrane proteins in order to investigate if the reduced glucose uptake rate of the mutants was beneficial for their production. The mutants were able to produce three out of five integral membrane proteins that were not possible to produce by the wild-type strain. The expression level of one selected membrane protein was increased when using the mutants and the expression level appeared to be a function of strain, glucose uptake rate and acetic acid accumulation. For production purposes, it is not uncommon that the recombinant proteins are secreted to the E. coli periplasm. However, one drawback with secretion is the undesired leakage of periplasmic products to the medium. The leakage of the product to the medium was studied as a function of the feed rate of glucose in fed-batch cultivations and they were found to correlate. It was also shown that the amount of outer membrane proteins was affected by the feed rate of glucose and by secretion of a recombinant product to the periplasm. The cell surface is another compartment where recombinant proteins can be expressed. Surface display of proteins is a potentially attractive production strategy since it offers a simple purification scheme and possibilities for on-cell protein characterisation, and may in some cases also be the only viable option. The AIDA-autotransporter was applied for surface display of the Z domain of staphylococcal protein A under control of the aidA promoter. Z was expressed in an active form and was accessible to the medium. Expression was favoured by growth in minimal medium and it seemed likely that expression was higher at higher feed rates of glucose during fed-batch cultivation. A repetitive batch process was developed, where relatively high cell densities were achieved whilst maintaining a high expression level of Z. / QC 20110608

Les autotransporteurs monomériques, appartenant au système de sécrétion de type V, correspondent à une famille importante de facteurs de virulence bactériens. Plusieurs fonctions, souvent essentielles pour le développement d’une infection ou pour le maintien et la survie des bactéries dans l’organisme hôte, ont été décrites pour cette famille de protéines. Malgré l’importance de ces protéines, notre connaissance de leur biogenèse et de leur mécanisme d’action demeure relativement limitée. L’autotransporteur AIDA-I, retrouvé chez diverses souches d’Escherichia coli, est un autotransporter multifonctionnel typique impliqué dans l’adhésion et l’invasion cellulaire ainsi que dans la formation de biofilm et d’agrégats bactériens. Les domaines extracellulaires d’autotransporteurs monomériques sont responsables de la fonctionnalité et possèdent pratiquement tous une structure caractéristique d’hélice β. Nous avons mené une étude de mutagenèse aléatoire avec AIDA-I afin de comprendre la base de la multifonctionnalité de cette protéine. Par cette approche, nous avons démontré que les domaines passagers de certains autotransporteurs possèdent une organisation modulaire, ce qui signifie qu’ils sont construits sous la forme de modules fonctionnels. Les domaines passagers d’autotransporteurs peuvent être clivés et relâchés dans le milieu extracellulaire. Toutefois, malgré la diversité des mécanismes de clivage existants, plusieurs protéines, telles qu’AIDA-I, sont clivées par un mécanisme qui demeure inconnu. En effectuant une renaturation in vitro d’AIDA-I, couplée avec une approche de mutagenèse dirigée, nous avons démontré que cette protéine se clive par un mécanisme autocatalytique qui implique deux acides aminés possédant un groupement carboxyle. Ces résultats ont permis la description d’un nouveau mécanisme de clivage pour la famille des autotransporteurs monomériques. Une des particularités d’AIDA-I est sa glycosylation par une heptosyltransférase spécifique nommée Aah. La glycosylation est un concept plutôt récent chez les bactéries et pour l’instant, très peu de protéines ont été décrites comme glycosylées chez E. coli. Nous avons démontré que Aah est le prototype pour une nouvelle famille de glycosyltransférases bactériennes retrouvées chez diverses espèces de protéobactéries. La glycosylation d’AIDA-I est une modification cytoplasmique et post-traductionnelle. De plus, Aah ne reconnaît pas une séquence primaire, mais plutôt un motif structural. Ces observations sont uniques chez les bactéries et permettent d’élargir nos connaissances sur la glycosylation chez les procaryotes. La glycosylation par Aah est essentielle pour la conformation d’AIDA-I et par conséquent pour sa capacité de permettre l’adhésion. Puisque plusieurs homologues d’Aah sont retrouvés à proximité d’autotransporteurs monomériques putatifs, cette famille de glycosyltranférases pourrait être importante, sinon essentielle, pour la biogenèse et/ou la fonction de nombreux autotransporteurs. En conclusion, les résultats présentés dans cette thèse apportent de nouvelles informations et permettent une meilleure compréhension de la biogenèse d’une des plus importantes familles de protéines sécrétées chez les bactéries Gram négatif. / Monomeric autotransporters, a family of proteins that use the type V secretion pathway, are important mediators of virulence for many bacterial pathogens. Many functions important for host colonization and survival have been described for these proteins. Despite the recognized importance of this family of proteins, the mechanisms that are required for the biogenesis and functionality of monomeric autotransporters still remain poorly understood. The Escherichia coli adhesin involved in diffuse adherence (AIDA-I) is a classical multifunctional autotransporter protein that mediates bacterial aggregation and biofilm formation, as well as adhesion and invasion of cultured epithelial cells. Extracellular domains of autotransporters are responsible for the protein function and fold into a characteristic β-helical structure. We performed a random mutagenesis of the AIDA-I passenger domain in order to identify regions involved in the various phenotypes associated with the expression of this protein. Our study suggests that the passenger domain of AIDA-I possesses a modular organization, which means that AIDA-I is built with individual functional modules. Autotransporter passenger domains can be cleaved from the β-domain and released into the extracellular milieu. However, despite the fact that diverse cleavage mechanisms have been previously described, many autotransporters, like AIDA-I, are cleaved by an unknown mechanism. By monitoring the in vitro refolding and cleavage following by site-directed mutagenesis, we showed that AIDA-I processing is an autocatalytic event that involves two acidic residues. Our results unveil a new mechanism of auto-processing in the autotransporter family. AIDA-I is one of the few glycosylated proteins found in Escherichia coli. Glycosylation is mediated by a specific heptosyltransferase encoded by the aah gene, but little is known about the role of this modification and the mechanism involved. Our findings suggest that Aah represents the prototype of a new large family of bacterial protein O-glycosyltransferases that modify various substrates recognized through a structural motif. Furthermore, we showed that glycosylation occurs in the cytoplasm by a cotranslational mechanism. These observations are unique in bacteria and represent a significant advance in our comprehension of prokaryotic glycosylation. We also showed that glycosylation is required to ensure a normal conformation of AIDA-I and, as a consequence, is necessary for its cell-binding function. The finding that other autotransporters or large adhesin-encoding genes are linked to Aah homologue-encoding genes suggests that glycosylation may be important, if not essential, for the function of these proteins, as for AIDA-I. In conclusion, the results presented in this thesis bring new information about the autotransporter family and also give new insight into the mechanisms that are important for different aspects of the biogenesis of monomeric autotransporters.