Abstract Bone sialoprotein (BSP) is a major component of medullary bone in chicken (Gallus gallus) and plays an important role in bone remodeling and mineralization. To explore the regulatory mechanism of BSP gene expression and the effect of variations in the promoter region on gene expression in laying hens,Hyline Grey laying hens were used as experimental animals,and the sequence variation in BSP promoter region was analyzed with gene cloning and sequencing technology. The online software JASPAR2018 was used for binding site prediction of transcription factors. Luciferase reporter gene recombinant vectors with different length of promoter fragments were constructed and transfected into chicken fibroblast line DF-1 whose dual-luciferase activity was measured. The expression of BSP gene was determined by qRT-PCR method. The results showed that there were 6 haplotypes (Hap1~6) in BSP promoter (-1257~+142 bp),and the core promoter region was located at -202~+142 bp. The difference of transcriptional activity was also found between Hap1 and Hap5 in the promoter region. The high transcriptional activity of Hap1 in the core promoter region caused an increase in the expression level of BSP gene. In conclusion,the core region in promoter of chicken BSP gene was identified and cloned successfully,which could provide basic materials for the investigation of regulatory mechanism of BSP gene expression and genetic selection of bone quality in laying hens.
WANG Han,YUE Qiao-Xian,ZHOU Rong-Yan, et al. Identification and Analysis of BSP Promoter in Laying Hens (Gallus gallus)[J]. 农业生物技术学报, 2020, 28(11): 2023-2031.
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