转录因子Sp1对成肌细胞中猪SKIP的表达调控作用

Role of Transcription Factors Sp1 in the Expressional Regulation of SKIP Gene in Porcine C2C12 Myoblasts

利用基因组PCR步移方法获得猪SKIP转录起始位点上游2 075 bp启动子序列。生物信息学分析发现该序列中存在4个Sp1结合位点和1个CpG岛。将启动子序列构建到pGL3-basic的双荧光素酶报告基因载体上,再利用RNA干扰技术分析转录因子Sp1对SKIP转录的影响。荧光素酶活性分析发现Sp1的表达抑制使成肌细胞中SKIP启动子活性显著下降,表明转录因子Sp1可能通过顺式作用元件GC-Box对成肌细胞分化过程中SKIP基因的转录激活起正向调控作用。

Genome walking technique was used to clone the proximal promoter region of porcine SKIP.A fragment of 2 075 bp upstream start code was obtained from genomic DNA of porcine.Bioinformatics analysis revealed that it had 4 Sp1 binding sites and one CpG island.The cloned SKIP promoter were cloned into the upstream of pGL3-basic to analyze the effect of Sp1 on SKIP transcription activity by RNAi technique.Luciferase activity analysis indicated a decrease in SKIP transcriptional activity through inhibiting the expression of transcription factor Sp1 in differentiating C2C12 myoblasts.These results suggested that the transcription factor Sp1 played a positive regulatory role in SKIP expression possibly by GC elements in myotubes.