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DNA methylation analysis of promoters in chicken brain by means of high resolution melting
Linköpings universitet, Institutionen för fysik, kemi och biologi, Zoologi.
2010 (engelsk)Independent thesis Advanced level (professional degree), 20 poäng / 30 hpOppgave
Abstract [en]

Domesticated and non­domesticated chickens differ in a range of morphological and behavioural traits, including differences in brain gene expression. DNA methylation of CpG dinucleotides in promoters is an important mechanism of epigenetic gene regulation. Previously, gene expression microarrays have been used to find differentially expressed genes in the brain of red junglefowl (RJF) and White Leghorn (WL) chickens. To investigate whether differential promoter DNA methylation can explain some of these expression differences, methylation sensitive high resolution melting (MS­-HRM) was used on five candidate genes taken from the expression data. Candidate genes were screened with bioinformatic software to find CpG islands overlapping predicted promoters. DNA was isolated from frozen optic tecta from a subset of the expression study birds. Bisulfite converted DNA was amplified with real­time PCR, melted, and compared to mixtures of methylated and non­methylated DNA. CpG islands adjacent to the differentially expressed transcript JMJD1C and an unannotated transcript on chromosome three were unmethylated, while a CpG island adjacent to TMEM208 was completely methylated. Additionally, CpG islands of ZMYND11 and LOC424014 appears to be at least 50% methylated, but these results are more insecure. Part of the β­-globin insulator, previously known to be methylated in non­expressing tissues, was completely methylated. The results indicate that this MS­-HRM method is suitable for methylation analysis in this material. No differential methylation between WL and RJF was found. For the unannotated transcript, JMJD1C and TMEM208, differential methylation as an explanation of differential expression in optic tecta can be excluded.

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2010. , s. -
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Identifikatorer
URN: urn:nbn:se:liu:diva-57706ISRN: LITH-IFM-A-EX-10/2291--SEOAI: oai:DiVA.org:liu-57706DiVA, id: diva2:327501
Presentation
2010-06-07, Linköpings universitet, 14:15 (engelsk)
Uppsök
Life Earth Science
Veileder
Examiner
Tilgjengelig fra: 2010-06-29 Laget: 2010-06-29 Sist oppdatert: 2011-03-24bibliografisk kontrollert

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