DNA methylation analysis of promoters in chicken brain by means of high resolution melting
Independent thesis Advanced level (professional degree), 20 credits / 30 HE creditsStudent thesis
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.
Place, publisher, year, edition, pages
2010. , - p.
IdentifiersURN: urn:nbn:se:liu:diva-57706ISRN: LITH-IFM-A-EX-10/2291--SEOAI: oai:DiVA.org:liu-57706DiVA: diva2:327501
2010-06-07, Linköpings universitet, 14:15 (English)
UppsokLife Earth Science
Jensen, Per, Professor
Jensen, Per, Professor