CAJ | 학술논문

前期通过染色质免疲沉淀技术(chromatin immunoprecipitation assay,CHIP)筛选出了转录因子activator protein-2 alpha(AP-2α)的靶基因Sumf1.采用重叠延伸PCR定点诱变技术,对AP-2α在Sumf1内含子片段上的两个结合位点的碱基进行定点突变,并构建定点突变表达载体.DNA测序表明,Sumf1内含子片段103～111 bp.411～419 bp两处的碱基已分别由GGGGTCAGG突变为GAAGTCCTG,由GCCTCTAGG突变为GGATCTCTG,成功实现定点诱变,为进一步研究AP-2α对基因Sumf1的表达调控奠定了基础.
전기통과염색질면피침정기술(chromatin immunoprecipitation assay,CHIP)사선출료전록인자activator protein-2 alpha(AP-2α)적파기인Sumf1.채용중첩연신PCR정점유변기술,대AP-2α재Sumf1내함자편단상적량개결합위점적감기진행정점돌변,병구건정점돌변표체재체.DNA측서표명,Sumf1내함자편단103～111 bp.411～419 bp량처적감기이분별유GGGGTCAGG돌변위GAAGTCCTG,유GCCTCTAGG돌변위GGATCTCTG,성공실현정점유변,위진일보연구AP-2α대기인Sumf1적표체조공전정료기출.
Through ChIP experiment, Sumf1 was screened out as one of the target genes of transcription factor activator protein-2 alpha(AP-2α). Site-directed mutagenesis method based on overlap extension PCR was used to introduce mutations in the two sites which were AP-2α binding sites in the Sumf1 intron fragment, and the site-directed mutagenesis eukaryotic expression vector were constructed. DNA sequencing showed that GGGGTCAGG of 103～111 bp sites had been changed into GAAGTCCTG and GCCTCTAGG of 411～419 bp sites had been changed into GGATCTCTG from mutagenesis. Site-directed mutagenesis was successfully implemented and laid the foundation for further study of AP-2α regulation of the expression of Sumf1.