鸡不同生精细胞中Piwil1基因启动子区DNA甲基化差异研究

doi:10.11843/j.issn.0366-6964.2014.09.004

Abstract

Abstract:

Transcription level and methylation in promoter region of Piwil1 gene in different types of spermatogenic cells were conducted to explore the mechanism of transcriptional regulation of Piwil1 gene in the process of spermatogenesis.By using fluorescence quantitative PCR technique to detect Piwil1 genes in different cells of mRNA expression level，and using Bisulfite sequencing PCR method for analysis of Piwil1 gene promoter region methylation，which included in modification，transformation，cloning and sequencing the promoter region.The results showed that Piwil1 gene expression of chicken was lowest in sperm，and the expression of spermatogonia was significantly higher than PGCs，SSCs and tetraploid(P＜0.01).And there was a CpG island founded in Piwil1 gene promoter（-233-+298 bp），which present 56 CpG sites.In the region of -233--129 bp(1-10 CpG sites)，the methylation sites were not founded in sperm cells，tetraploid cells，PGCs and SSCs，while the methylation ratio of spermatogonia was up to 0.600; however，in the region of -233--129 bp(39-55 CpG sites)，significant differences in methylation ratio between different cells were detected，in which the methylation ratio of spermatogonia was the lowest to 0.212，while PGCs and SSCs in DNA hypermethylation inhibited the expression of genes Piwil1 in a certain degree.It was concluded that methylation in +105-+252 bp promoter regions may have some influence on transcription regulation of Piwil1 gene，but there were other transcriptional regulatory mechanisms.

CLC Number:

S831.2

CHEN Jing，ZHAI Fei，XIA Ming-xiu，CHEN Rong，XU Qi，CHANG Guo-bin，LI Jian-chao，MA Teng，WANG Hong-zhi，XU Lu，LIU Lu，CHEN Guo-hong. Differences in DNA Methylation of the Promoter Region of Piwil1 Gene in Chicken Spermatogenic Cells[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(9): 1404-1409.

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Differences in DNA Methylation of the Promoter Region of Piwil1 Gene in Chicken Spermatogenic Cells

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