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183 Uppsatser om Reporter gene assay - Sida 1 av 13

?-galaktosidas assay för studie av promotorregion i kloritdismutas från Ideonella dechloratans

Oxochlorates are anions with a partially naturally occurrence in nature but are also spread by human activities, including the paper industry. These compounds are harmful to both nature and humans, which makes it necessary to find a good way for their degradation. There are two different kinds of bacteria that can use oxochlorates as electron acceptors in their metabolism, bacteria that break down perchlorate and bacteria that break down both perchlorate and chlorate. A bacterium that can break down chlorate under anaerobic conditions is Ideonella dechloratans which holds the genes for chlorite dismutase and chlorate reductase which are enzymes for the degradation of chlorate. Gene expression and enzyme activity of chlorite dismutase are induced under anaerobic conditions, which makes it interesting to find out how this regulation functions in order to better exploit these bacteria in biological wastewater treatment.

Kloning av möjlig promotorsekvens uppströms kloritdismutas i Ideonella dechloratans.

Perchlorate and chlorate are naturally occurring in the atmosphere, from here it sediments into groundwater and soil. The pollution is increased by discharges of perchlorate and chlorate from agriculture and paper mills. Bacteria capable of reducing perchlorate and chlorate to chloride and oxygen can be used to get rid of these contaminants. However an anaerobic environment needs to be sustained in order for this reaction to be used. For this reduction to work in an aerobic environment as well, a greater knowledge of the reducing enzymes, regulating factors and their corresponding genes is needed.

VERIFIERING AV ANALYS FÖR KONCENTRATIONSMÄTNING AV BIOLOGISKA LÄKEMEDEL

Biologiska läkemedel är stora molekyler som har renats eller producerats från ett biologiskt ursprung. De används idag i stor utsträckning runt om i världen på grund av deras effektivitet att minska symtomen som kan uppstå vid olika mycket svåra sjukdomar. Detta arbete fokuserar på biologiska läkemedel som hämmar cytokinet tumörnekrotisk faktor (TNF). Läkemedlen används bland annat vid svåra autoimmuna sjukdomar. Vid autoimmuna sjukdomar aktiveras cellerna i immunsystemet vilket leder till utsöndring av olika cytokiner till exempel interleukin 1 och TNF.

VERIFIERING AV ANALYS FÖR KONCENTRATIONSMÄTNING AV BIOLOGISKA LÄKEMEDEL

Biologiska läkemedel är stora molekyler som har renats eller producerats från ett biologiskt ursprung. De används idag i stor utsträckning runt om i världen på grund av deras effektivitet att minska symtomen som kan uppstå vid olika mycket svåra sjukdomar. Detta arbete fokuserar på biologiska läkemedel som hämmar cytokinet tumörnekrotisk faktor (TNF). Läkemedlen används bland annat vid svåra autoimmuna sjukdomar. Vid autoimmuna sjukdomar aktiveras cellerna i immunsystemet vilket leder till utsöndring av olika cytokiner till exempel interleukin 1 och TNF.

The WT1-gene ? its role in tumourigenesis and prospects for developing a vaccine

The WT1 gene is a complex gene originally known to suppress cancer in kidneys. Studies of WT1 knockout mice have confirmed the important role of WT1 in the pathogenesis of Wilms? tumour, a tumour which counts for 95% of all childhood renal tumours. In that case the WT1 gene acts as a tumour suppressor gene. Subsequent research has shown that the WT1 gene in many other cases acts as an oncogene, e g in leukemia or lung cancer (even though these cancer forms can emerge as a result of many other aetiological factors).

The RECK gene and invasive cancer development : the significance of RECK in angiogenesis and inhibition of matrix metalloproteinases

The RECK gene is a relatively new discovered gene with important implications for cancer research. The research has been primarily concentrated on the human gene with the ultimate aim to identify the invasive characteristics. Up regulated RECK is linked to significantly prolonged survival rates in patients with severe forms of malignancies. RECK is normally expressed in all cells of the body and has an important role in the balance between destructive and constructive features of the extracellular matrix. The RECK protein is a membrane-bound glycoprotein that inhibit matrix metalloproteinases which has the function of breaking down the ECM. There is a significant correlation between RECK gene expression and the formation of new vessels, presumably via the mediation of VEGF which is an important and powerful inducer of angiogenesis.

Gene expression from a cold-treated Swedish isolate of Haemonchus contortus

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.

Validation of a tetraplex assay for detection of antibodies in poultry serum using Luminex 200 platform.

Background: As a part of a national health control program, Statens VeterinärmedicinskaAnstalt performs diagnostics to screen flocks for certain pathogens causing high mortality,morbidity and/or serious economical losses. There are several viruses in the programincluding IBDV (infectious bursal disease virus), IBV (infectious bronchitis virus) and NDV(Newcastle disease virus). Method: 96 serum samples were collected from different poultryflocks in Sweden and analyzed by ELISA, which are currently used in the health controlprogram as well as by a commercial prototype of a multiplex immunoassay manufactured byLuminex Corp., which is currently under evaluation at the United States Department ofAgriculture USDA. This 4-plex assay detects antibodies for the three above-mentionedviruses as well as antibodies of avian reovirus. In the context of this study the ELISAs run inroutine diagnostics as well as a REO ELISA were used as the standard for comparison.Result: The antibody concentration in serum from vaccinated chickens was high while theantibody concentration level in serum from not vaccinated chickens was low.

Minor fieled study - molecular epidemiologi of rabies in Sao Paulo state and Minas Gerais state, Brazil

Rabies is a fatal zoonotic disease spread worldwide. The most common sources of infection for all animals and humans are bites from dogs or bats. The aim of this degree project was to diagnose and determine the source of infection for 11 rabies samples from São Paulo State and Minas Gerais State, Brazil. Diagnosis was made through direct immunofluorescence assay, mouse inoculation and RT-PCR. The sources of infection were determined by sequencing 234 nucleotides of the 5? end of the N-gene and align these with homologous sequences retrieved from GenBank.

Species diversity and geographical distribution of Fusarium species on winter wheat in two regions in Sweden

Various Fusarium species causes some of the most common cereal diseases worldwide. Besides the yield losses that can be a result of these diseases, strains from several Fusarium species can produce mycotoxins, some very toxic. The aim of this study was to investigate which Fusarium species and how many that occurred in winter wheat grains from Mälardalen and Kalmar län, if there was any difference in the distribution of Fusarium fungi between the regions and the potential within species to produce mycotoxins belonging to the group trichothecenes. Winter wheat grains collected in 2009 from ten fields (unsprayed plots) in the two regions were analysed for Fusarium species. PCR was used to amplify the TEF region where after the samples were sequenced.

The screening for novel proteasome inhibitors as a treatment of cancer using IncuCyte FLR and fluorometric microculture cytotoxicity assay.

The problem of finding targeted medicine is a central problem in chemotherapy. From this point of view the ubiquitin-proteasome system is a highly promising object in the pharmaceutical approach. Proteasome plays a critical role in cellular protein degradation, cell cycle and apoptosis regulation.Proteasome inhibitors are substances blocking the actions of proteasome. Cancer cells are more sensitive to inhibition of the ubiquitin-proteasome system than normal cells. Therefore proteasome inhibitors have the potential to be successfully used in the cancer treatment.The study aimed to test various substances to identify possible proteasome inhibitors with the IncuCyteTM FLR image system and fluorometric microculture cytotoxicity assay.

Evaluation of an automated multiplex real-time RT-PCR assay for rapid detection of Influenza A and B viruses

Influenza is a viral infection that affects global health and economy with its endemic and sometimes pandemic spread. Rapid detection of Influenza viruses enables antiviral use and can bring financial savings. It is also essential for the global surveillance of prevalent Influenza strains. RT-PCR is considered the most specific and sensitive method for detection of Influenza, but Influenza mutates at a high rate and it is therefore crucial that RT-PCR methods are updated regularly.In 2014, Cepheid released their Xpert Flu/RSV XC assay, which can detect Influenza A and B and RSV by multiplex RT-PCR in approximately one hour. The aim of this study was to evaluate this assay at Laboratoriemedicin Västernorrland by using the laboratory?s previous PCR assay for detection of Influenza viruses as reference method.Real-time RT-PCR was used to compare Xpert Flu/RSV XC to the reference method.

Optimization and validation of a triplex real-time PCR assay for thermotolerant Campylobacter species associated with foodborne disease

The genus Campylobacter is globally recognised as the leading bacterial cause of human foodborne gastroenteritis. Every year around 8000 Swedes are infected by Campylobacter. Most people are infected by thermotolerant Campylobacter species, commonly C. jejuni and C. coli.

Optimization of immunoassay parameters in multiplex in the high throughput protein detection technique Proximity Extension Assay

The ability to detect protein-based biomarkers, which are linked to different diseases like colorectal cancer, is very important as a diagnostic tool. Usually complex biological samples like blood are studied which will contribute to different technical issues when performing an assay. The aim with the project is to optimize and develop the high throughput protein detection technique Proximity Extension Assay, PEA, into a 96-plex panel, in hopes of discovering an expression profile for colorectal cancer. PEA was developed by Olink Bioscience and allows specific proteins in a sample to be quantitatively transformed into nucleic acid sequences that are subsequently detected and quantified with real-time PCR. Two proximity probes containing oligonucleotide sequences bind pairwise to target protein and when brought in proximity, a DNA polymerase will extend a hybridization arm from one probe over to the second forming a double-stranded DNA sequence that can serve as a template in real-time PCR.

Development of real-time RT-PCR for the detection of human sapovirus in foods

Food-poisoning is a major health problem and an estimated half a million Swedes are food-poisoned annually, with acute gastroenteritis as a consequence. One of the major causes of contaminated foods is related to food- and waterborne viruses. To be able to trace back the source of contaminant, the method of detecting viruses must be specific and sensitive. No standardized method for detecting foods for sapovirus exists today. The aim of the work described in this bachelor thesis is to implement and opti-mize a real-time RT-PCR method for the detection of all genogroups of human sapovirus in foods.

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