Metodutveckling för analys av serglycinuttrycket i blodet hos hundar
The development of quantitative, real-time PCR (qPCR) combined with the mapping of the canine genome opens new possibilities in veterinary medicine. This method provides a quick and accurate quantification of the expression of a specific gene at a given point in time and thereby also information of...
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| Formato: | L3 |
| Lenguaje: | sueco Inglés |
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SLU/Dept. of Anatomy, Physiology and Biochemistry (until 231231)
2008
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| Materias: |
| _version_ | 1855571996640804864 |
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| author | Sandros, Boel |
| author_browse | Sandros, Boel |
| author_facet | Sandros, Boel |
| author_sort | Sandros, Boel |
| collection | Epsilon Archive for Student Projects |
| description | The development of quantitative, real-time PCR (qPCR) combined with the mapping of the canine genome opens new possibilities in veterinary medicine. This method provides a quick and accurate quantification of the expression of a specific gene at a given point in time and thereby also information of how the gene expression for a certain protein is influenced by various conditions and diseases. One possible area of application is identifying bio markers for cancer. Recently the protein serglycin (the core protein of an intracellular proteoglycan) was found to act as a selective marker for the disease acute myeloid leukemia in humans (Niemann et al, 2007). Serglycin is produced by most hematopoietic cells, although mast cells account for the largest amount of serglycin. The expression of serglycin in dogs remains to be explored. As mast cell tumours are common in this species, it would be of interest to find out whether serglycin can be used as a marker for these tumours, to facilitate diagnosis and prognosis as well as evaluation of how well a patient is responding to treatment.
The aim of this study is to develop a qPCR-analysis of serglycin expression in canine blood, to serve as a starting point for further research. Several parameters, such as the optimum temperature for the PCR-reaction, primer design and DNA-concentration, have been established. The resulting PCR-analysis is functioning, although still in need of further modifications in order to achieve the desired efficiency and reproducibility. |
| format | L3 |
| id | RepoSLU12083 |
| institution | Swedish University of Agricultural Sciences |
| language | swe Inglés |
| publishDate | 2008 |
| publishDateSort | 2008 |
| publisher | SLU/Dept. of Anatomy, Physiology and Biochemistry (until 231231) |
| publisherStr | SLU/Dept. of Anatomy, Physiology and Biochemistry (until 231231) |
| record_format | eprints |
| spelling | RepoSLU120832017-11-08T08:47:02Z Metodutveckling för analys av serglycinuttrycket i blodet hos hundar Sandros, Boel serglycin realtime PCR kvantitativ PCR RT PCR mRNA The development of quantitative, real-time PCR (qPCR) combined with the mapping of the canine genome opens new possibilities in veterinary medicine. This method provides a quick and accurate quantification of the expression of a specific gene at a given point in time and thereby also information of how the gene expression for a certain protein is influenced by various conditions and diseases. One possible area of application is identifying bio markers for cancer. Recently the protein serglycin (the core protein of an intracellular proteoglycan) was found to act as a selective marker for the disease acute myeloid leukemia in humans (Niemann et al, 2007). Serglycin is produced by most hematopoietic cells, although mast cells account for the largest amount of serglycin. The expression of serglycin in dogs remains to be explored. As mast cell tumours are common in this species, it would be of interest to find out whether serglycin can be used as a marker for these tumours, to facilitate diagnosis and prognosis as well as evaluation of how well a patient is responding to treatment. The aim of this study is to develop a qPCR-analysis of serglycin expression in canine blood, to serve as a starting point for further research. Several parameters, such as the optimum temperature for the PCR-reaction, primer design and DNA-concentration, have been established. The resulting PCR-analysis is functioning, although still in need of further modifications in order to achieve the desired efficiency and reproducibility. Utvecklandet av kvantitativ, direktanalyserad PCR (qPCR, realtidsPCR) öppnar tillsammans med kartläggningen av hundens genom nya möjligheter inom veterinärmedicinen. Med denna metod kan man snabbt och noggrant kvantifiera uttrycket av en specifik gen och på så vis få information om hur genuttrycket för ett visst protein påverkas vid olika tillstånd. Ett möjligt användningsområde är att identifiera biomarkörer för olika cancersjukdomar. Nyligen konstaterades proteinet serglycin (som utgör kärnan i en intracellullär proteoglykan) vara en selektiv markör för sjukdomen akut myeloid leukemi hos människa (Niemann et al, 2007). Serglycin förekommer hos flertalet hematopoietiska celler, dock svarar mastcellerna för den högsta produktionen. Till min vetskap har serglycinuttrycket hos hund ännu ej studerats. Eftersom mastcellstumörer är vanliga hos detta djurslag vore det intressant att ta reda på om serglycin kan användas som biomarkör för dessa, att användas såväl prognostiskt och diagnostiskt som för uppföljning av insatt behandling. Syftet med detta arbete är att utveckla en qPCR-analys av serglycinuttryck i hundblod som kan ligga till grund för fortsatta studier. Olika parametrar, såsom optimal temperatur för PCR-reaktionen, primerdesign och DNA-koncentration, har fastslagits. Slutresultatet blev en fungerande qPCR-analys för hundserglycin, som dock behöver ytterligare modifieringar för att uppnå önskad effektivitet och reproducerbarhet. SLU/Dept. of Anatomy, Physiology and Biochemistry (until 231231) 2008 L3 swe eng https://stud.epsilon.slu.se/12083/ |
| spellingShingle | serglycin realtime PCR kvantitativ PCR RT PCR mRNA Sandros, Boel Metodutveckling för analys av serglycinuttrycket i blodet hos hundar |
| title | Metodutveckling för analys av serglycinuttrycket i blodet hos hundar |
| title_full | Metodutveckling för analys av serglycinuttrycket i blodet hos hundar |
| title_fullStr | Metodutveckling för analys av serglycinuttrycket i blodet hos hundar |
| title_full_unstemmed | Metodutveckling för analys av serglycinuttrycket i blodet hos hundar |
| title_short | Metodutveckling för analys av serglycinuttrycket i blodet hos hundar |
| title_sort | metodutveckling för analys av serglycinuttrycket i blodet hos hundar |
| topic | serglycin realtime PCR kvantitativ PCR RT PCR mRNA |