Abstract:

【Objective】 The fatty acid synthase (FAS) gene promoter of Xinong Saanen dairy goat was cloned and sequenced to analyze the active region, thus providing an evidence for function determination and expression regulation of FAS gene. 【Method】 According to the homologous sequences of bovine and homo, and to 5‘UTR of Xinong Saanen dairy goat FAS gene, upstream and downstream primers were designed, respectively. Using genome DNA as the template, the FAS gene promoter was cloned. Primers were redesigned according to the results of bioinformatics analysis, the FAS gene promoter was cloned by segmenting and linking up with the PGL-3 expression vector, co-transfecting 293 and MCF-7 cells with the β-galactosidase control vector, and luciferase and β-galactosidase expression were detected. 【Result】 There are 2 640 bp in FAS gene promoter totally, the homology with bovine and homo is over 90%. The promoter contains several potential transcription factor bind sites such as SP1, Ets, LSF bind sites, several CCAAT boxes and a GC box. The core promoter sequences potentially locate in -1 040—-340 bp, and reduced the range to -721—-540 bp by promoter deletion experiment. 【Conclusion】 The full length of Xinong Saanen Dairy Goat FAS gene promoter was cloned, the potential negative control element was analyzed, and the core promoter sequences were found.

Key words: Xinong Saanen dairy goat, FAS promoter, gene cloning, activity determination