Effect of c.36-2277T>C mutation on the expression of luciferase in chicken GNAS gene

• Published in: 浙江大学学报. 农业与生命科学版
• Main Authors: WANG Huanhuan, ZHANG Lei, GE Ying, SONG Dandan, LOU Lifeng, ZHANG Xuedong
• Format: Article
• Language: English
• Published: Zhejiang University Press 2019-04-01
• Subjects: chicken; <italic>GNAS</italic> gene; mutation; luciferase
• Online Access: https://www.academax.com/doi/10.3785/j.issn.1008-9209.2018.03.131
• ISSN: 1008-9209; 2097-5155

In order to analyze the effect of ENSGALT00000062075.1:c.36-2277T &gt; C mutation on the expression of transcriptional regulation in chicken GNAS gene, this study collected blood DNA samples from black-bone chicken and barred chicken to construct different genotypic plasmids of c. 36-2277T&gt;C mutation, and detected the expression of dual-luciferase reporter analysis system after transfecting chicken embryo fibroblasts. The result showed that all barred chicken samples belonged to TT genotypes and all black-bone chicken samples were CC genotypes. The section from 175 bp upstream to 818 bp downstream of c.36-2277T&gt; C mutation had significantly transcript enhancing activity (P&lt;0.01). The activity of CC genotype was significantly higher than that of the TT genotype (P&lt;0.000 1). The result of transcript factor prediction showed that the number of TGGCA-binding protein sites was the largest in this 993 bp section, and the adjacent mutations were AP-1 and ER-alpha/T3R-alpha binding sites. The study suggests that the base types of c.36-2277T&gt;C mutation in GNAS gene have an important impact on the process of gene transcription regulation, and this mutation plays the role associated with upstream and downstream binding sites of transcript factors.