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

作　　者：熊琪[1] 柴进[2] 张年[1] 索效军[1] 李晓锋[1] 杨前平[1] 刘洋[1] 陈明新[1] 

机构地区：[1]湖北省农业科学院畜牧兽医研究所/动物胚胎工程及分子育种湖北省重点实验室,武汉430064 [2]华中农业大学动物科技学院/农业动物遗传育种与繁殖教育部重点实验室,武汉430070

出　　处：《湖北农业科学》2012年第18期4147-4151,共5页Hubei Agricultural Sciences

基　　金：湖北省动物胚胎工程及分子育种重点实验室开放课题(2010ZD111);湖北省科技计划自然科学基金重点项目(2010CBB01301);湖北省农业科学院青年科学基金(2011NKYJJ16);华中农业大学自主创新基金(2010BQ001);华中农业大学青年教师科技创新项目(2011QC001);湖北省农业科技创新中心(2011-620-001-003)

摘　　要：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.

关 键 词：SP1 猪 SKIP 启动子活性 RNA干扰 

分 类 号：Q789[生物学—分子生物学]