This review article presents the splicing process during messenger RNA maturation and how it is regulated by different Cis-regulatory RNA-sequence elements and splicing factors. A more detailed description of the process alternative splicing and its importance to the function of genes from the model organism Arabidopsis thaliana is also given. A single eukaryotic gene can by the process alternative splicing (AS) give rise to a number of functionally mature mRNA-molecules, which in turn encodes for structurally and/or functionally different proteins. During the course of evolution, the process alternative splicing has thus shown to be effective in increasing transcriptome and proteome diversity of most eukaryotic organisms. This suggests therefore that the dominant theory in molecular biology, a gene encodes for a protein, needs to be corrected.

Alternativ splicing av pre-mRNA ger upphov till proteindiversitet. Histonmodifieringar kopplas till den alternativa splicingens reglering genom adaptorsystem som overfor den epigenetiska informationen direkt till splicingfaktorerna. De cis- agerande RNA- elementen pa exoner och introner med tillhorande trans- reglerande splicingfaktorer paverkas darfor direkt av specifika histonmodifieringar. En sammankopplande integrerad modell over en rad DNA- baserade processer foreslas. Denna komplexa modell ger en bild av interaktioner och paverkan mellan dessa delar.

Exoner är de sekvenser i DNA vilka rymmer koden för proteiner i människan och i alla andra organismer. Intronerna, vilka utgör utrymmet mellan exoner, består av ickekodande sekvenser och kontrollelement. Exoner tillhörande en gen måste inte alltid inkluderas i den slutliga mRNA produkten, alternativ splicing tillåter exkludering av vissa sekvenser och gör att en gen kan ha mer än en mRNA produkt, därigenom kan en gen koda för flera olika proteiner. Alternativ splicing är ett fält som snabbt utvecklas och dess relevans för många sjukdomar har blivit uppenbar. Detta arbete går igenom ett flertal av dessa sjukdomar för att sammanställa ny forskning och tydliggöra rollen av alternativ splicing i dem.

Alternativ splicing är en process som ger upphov till att olika mRNA-sekvenser bildas från en enda gen, vilket bidrar till en ökad proteindiversitet hos organismen. Olika mRNA-sekvenser kan uppstå eftersom att det förekommer olika varianter av alternativ splicing som även kan kombineras på flera olika sätt: cassette exon (inkludering/exkludering av exon), intron retention (intronet behålls), alternative 5´splice-site choice (olika 5´ splice sites kan väljas) och slutligen alternative 3´ splice-site choice (andra 3´ splice sites kan väljas). För att alternativ splicing ska äga rum i olika pre-mRNA måste den regleras av cis-reglerande element. De cis-reglerande elementen utgörs av fyra grupper: exonic splicing enhancers (ESE), exonic splicing silencers (ESS), intronic splicing enhancers (ISE) samt intronic splicing silencers (ISS). Som namnen förtäljer finns de antingen i exoner eller introner, där de interagerar med transagerande faktorer, SR-proteiner (aktiverare) eller hnRNPs (hämmare).

Alternative splicing is a process that partly rejects the common definition of a gene ? that one gene codes for one specific protein. By variable combination of coding regions (exons) and exclusion of non-coding regions (introns), formation of several different mRNA-transcripts, and consequently several different proteins, can derive from the same gene. Alternative splicing is an important condition for the development of complex life forms, but it is also a highly sensitive process and inaccurate splicing is the cause of approximately 15 % of mutations that cause genetic diseases. This article presents four genes, BRCA1, BRCA2, ER? and ER?, and inaccurate splicing of these genes increases the risk of developing cancer, particularly breast cancer and ovarian cancer.

Abstract:In previous experiments with the bacterium Ideonella dechloratans a probable sequencecoding for a cytochrome c protein (cyt c) was observed, the sequence is assumed to have afunction in the bacterial respiratory chain. Detection attempts have been made to try to findthis protein, but have not succeeded. The purpose of this project is to detect the expressionof mRNA associated with the cytochrome c. And if it is detected, it will also be investigated ifthere is any difference in expression of the mRNA due to aerobic or anaerobic environment.Total RNA was purified from cultures grown under aerobic and anaerobic conditions. cDNAwas then synthesized using reverse transcriptase and subsequently amplified with thespecific primers in a qRT-PCR.qRT-PCR testing showed a distinct amplification of primer product, confirming that I.dechloratans expresses mRNA coding for the cyt c protein.

Tissue factor (TF), a 47 kDa glycoprotein, is the initiator of the extrinsic pathway of blood coagulation and consequently of the upmost importance when damage to blood vessel occurs. The source of TF in circulation has been investigated. However, the source of TF is still not clear. One theory is that platelets express and increases the expression of TF after stimulation and the aim of our report was to investigate whether platelets really are a source for TF in circulation.Using specific primers for TF mRNA, platelets in plasma from healthy volunteers and from patients suffering from cardiac infarction were analyzed by using real-time polymerase chain reaction (PCR). Gel electrophoresis was performed after amplification of TF mRNA to verify the results.The samples were negative for TF when using real-time PCR and the few positive all had cycle threshold (Ct) values above 35.

The Bovine respiratory syncytial virus (BRSV) causes bronchiolitis and interstitial pneumonia, predominantly in calves, and is a major cause of bovine respiratory disease worldwide. In humans, BRSV is paralleled by the closely related Human respiratory syncytial virus (HRSV), an important cause of respiratory disease, most severe in infants.The clinical signs and pathology during RSV infection is caused, not only by the direct effects of viral replication, but also by the response of the host immune system. The immunopathology of RSV has long obfuscated our understanding of the disease, and development of effective treatment and vaccines will be very difficult until greater knowledge is gained.One of the components of the immune system that has come into focus in RSV research the last few years, is the Toll-like receptor 4 (TLR4). The TLR4 receptor is well known as the receptor that binds lipopolysaccaride (LPS), and initiates the host response to bacterial infection. Recently, it has been shown that the fusion protein of RSV also interacts with, and up-regulates the expression of, the TLR4 receptor.

The purpose of this thesis is to analyze the variation in the maximal discrimination of the interaction between cognate and a non-cognate codon and anti-codon (also called the d-value). The variation was analyzed with a multiple regression model with the d-value as the dependent variable and with the codon position and the different mRNA and tRNA bases as independent variables. The result of the analysis not only confirmed earlier studies that the maximal accuracy was highest in the second codon position and lowest in the third codon position but we also found significant relationships and interaction effects..

The Bovine respiratory syncytial virus (BRSV) causes bronchiolitis and interstitial pneumonia, predominantly in calves, and is a major cause of bovine respiratory disease worldwide. In humans, BRSV is paralleled by the closely related Human respiratory syncytial virus (HRSV), an important cause of respiratory disease, most severe in infants.The clinical signs and pathology during RSV infection is caused, not only by the direct effects of viral replication, but also by the response of the host immune system. The immunopathology of RSV has long obfuscated our understanding of the disease, and development of effective treatment and vaccines will be very difficult until greater knowledge is gained.One of the components of the immune system that has come into focus in RSV research the last few years, is the Toll-like receptor 4 (TLR4). The TLR4 receptor is well known as the receptor that binds lipopolysaccaride (LPS), and initiates the host response to bacterial infection. Recently, it has been shown that the fusion protein of RSV also interacts with, and up-regulates the expression of, the TLR4 receptor.

Recurrent Airway Obstruction, RAO, är en icke infektiös luftvägssjukdom hos häst. Sjukdomen karaktäriseras av perioder med reversibel luftvägsobstruktion och inflammation orsakad av bronkospasm och ansamling av mucus och neutrofiler i luftvägarna. Sjukdomsorsaken anses vara multifaktoriell med ett samspel mellan dålig miljö och en viss genetisk predisposition. Den immunologiska bakgrunden till sjukdomen är fortfarande oklar men både den förvärvade och medfödda immuniteten anses delta i luftvägsinflammationen. Toll-lika receptorer, TLR, hör till de viktigaste receptorerna i den medfödda immuniteten.

Totally 84 differentially expressed mRNA clones from infective L3 larvae of the parasite Haemonchus contortus, a blood sucking nematode, were analyzed with single strand hybridization assay (SSH). Altogether 79 clones were sequenced, edited, and compared with proteins found via BLAST in GeneBank. The aim was to investigate gene expression and potential protein expression following storage at 5 °C for 32 weeks. mRNA was extracted from fresh and stored L3. The SSH derived products were cloned into E.

Inflammatory airway disease (IAD) is a respiratory disorder that most commonly affects young horses performing at a high level. The clinical signs are often mild but the disorder can cause a marked decrease in performance to a point where the horse has to be taken of training and competing. The aetiology underlying IAD is not yet fully understood. One current theory is that a type I-hypersensitivity reaction due to allergens in the environment is the cause in some horses. The most reliable diagnostic method is a bronchoalveolar lavage (BAL) since it reflects the inflammatory status in the most distal parts of the lungs, which are affected in IAD.

Immune stimulating complex (iscom) matrix is a formulation that is used as anadjuvant within research. The capability of the iscom matrix to elicit an immuneresponse when not present together with an antigen has not been extensivelyinvestigated. One way to evaluate the status of the immune system is to measure theamount of cytokines that usually are produced by various cell types during an immuneresponse. The aim of the present study was to evaluate the early immune responses inporcine peripheral blood mononuclear cells (PBMC) after stimulation in vitro with theiscom matrix, AbISCO®-100. The immune response was evaluated by measuring themRNA expression for various cytokines, a chemokine receptor (CXCR4) andregulator of G-protein signaling (RGS16) in porcine PBMC after stimulation withAbISCO®-100 or other inducers like a lipopolysaccharide (LPS) and a CpGoligodeoxynucleotide (ODN 2216) used as controls.

We where asked to write a webapplication, with similar functionalityas an existing desktop application. The application can todayrecord sounds from a person, mix it together at the optimal place withanother, similar sound and play back the result.The application used mathematical calculation that the developerswhere not familiar with, therefore it was decided to focus on the partsthat involved recording and playing sound, to pave the way for anyfuture implementation.The follwing prototypes where implemneted:1. Read a sound clip from a file.2. Splicing of two sound clips at different points in the clips.3. Generate the sound of a guitar.The result of this evaluation is that web browsers today are notready for this type of application, but that there is potetial as soonas W3C have standardised the use of a microphone as a source to theexisting DSP functions..