关键词: 牛, Sry, 核心启动子, 双荧光素酶报告基因分析系统

Abstract:

【Objective】 Sry is the pivotal gene initiating male sex determination in most mammals, but how its expression is regulated is still not clear. The study was to identify the regulation sequences of bovine Sry promoter, and to understand the transcriptional regulation of the Sry. 【Method】 A 1056 bp long bovine Sry 5'-flanking sequences, and the potential transcriptional start sites in this region were predicted by bioinformatics method as well as reporter gene analysis was isolated, Bovine genital ridge cells were isolated and deletion recombinant plasmids with different lengths of Sry 5' flanking regions were constructed. Sry 5'-flanking promoter activity analysis was performed in bovine genital ridge cells by the dual-luciferase reporter assay system.【Result】 The results indicated that the genital ridge cells cultured in vitro could express the characteristic genes of male genital ridge, including Sry, Sox9, Sf-1 and Dax1. Through the prediction analysis, three potential transcription start sites for bovine Sry were identified at -93, -419 and -722 upstream from the translation initiation site. Further analysis demonstrated that a 35 bp long fragment (-599 to -565) in the 5'-flanking sequence was important for the basic transcriptional activity. Prediction of the 35 bp long fragment demonstrated that there were several potential transcription factor binding sites in this region. 【Conclusion】 There were several potential regulation sequences existed in the 35 bp long fragment (-599 to -565) of bovine Sry 5'UTR。

Key words: Bovine, Sry, core promoter, dual-luciferase reporter assay system