蛋鸡骨钙素基因启动子变异及其转录活性研究

doi:10.3969/j.issn.1674-7968.2019.03.012

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Abstract Osteocalcin (OCN) is a kind of non-collagen bone matrix protein secreted and synthesized by osteoblasts, which plays an important role in bone metabolism such as bone remodeling and bone mineralization. This study aimed to investigate the mechanism of gene expression regulation of OCN in laying hens (Gallus gallus domesticus). Twenty 50-week-old DAWUFEN laying hens were selected and the blood was collected for DNA extraction. The primers for promoter region of OCN gene were designed according to the template sequence downloaded from Ensembl database (ENSGALG00000029494). Potential genetic variations in the promoter region of OCN gene were identified by Sanger sequencing. JASPAR2018 was used for transcription factors binding site prediction. The luciferase reporter gene recombinant vectors with different haplotypes were constructed and the dual-luciferase activity was detected by transfecting chicken fibroblast line DF-1. The results showed that the promoter region of the OCN gene in the amplified layer had the transcriptional activity and contained binding sites for bone metabolism-related transcription factors such as Vitamin D receptor (VDR), Runt related transcription factor (RUNX) 1, RUNX2, and RUNX3. Sequencing results revealed the presence of the insert fragment CCGCACTCTGCACTTTGCGGCCG and the deletion fragment CCACA in promoter region, as well as C>G and A>G variations. These variants constituted 4 haplotypes. The dual luciferase assay showed that all the 4 haplotypes in the promoter region had transcriptional activity, but the activity of different haplotypes had extremely significant difference (P<0.01), and the transcriptional activity of haplotype Ⅳ was the highest. In this study, the core promoter region of the OCN gene was identified, and there were differences in the promoter transcriptional activities of different haplotypes. A theoretical basis for the transcriptional regulation of OCN gene could be helpful for bone metabolism of laying hens.

Key words： Laying hen Bone metabolism Osteocalcin (OCN) Promoter

Received: 13 August 2018

ZTFLH:

S831.2

Corresponding Authors: * rongyanzhou@126.com

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	WANG Han
	CHEN Ye
	YUE Qiao-Xian
	ZHOU Rong-Yan
	CHEN Hui
	WANG De-He
	XI Jian-Zhong

Cite this article:

WANG Han,CHEN Ye,YUE Qiao-Xian, et al. Study on Variations and Transcriptional Activity of Osteocalcin Gene Promoter in laying hens (Gallus gallus domesticus)[J]. 农业生物技术学报, 2019, 27(3): 481-487.

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http://journal05.magtech.org.cn/Jwk_ny/EN/10.3969/j.issn.1674-7968.2019.03.012 OR http://journal05.magtech.org.cn/Jwk_ny/EN/Y2019/V27/I3/481

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