生命科学研究2009，Vol.13Issue(5)：430-436,7.
重叠延伸PCR法构建小鼠Sumf1基因的定点突变真核表达载体
Site-directed Mutagenesis of Sumf 1 Gene of Mouse Based on Overlap Extension PCR and Construction of Eukaryotic Expression Vector
摘要
Abstract
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.关键词
转录因子AP-2α/硫酸酯酶修饰因子Sumf1/重叠延伸PCR/定点诱变Key words
transcription factor AP-2α/ Sulfatase-modifying factor 1 Sumf1/ overlap extension PCR/ site-directed mutagenesis分类
生物科学引用本文复制引用
帅勇,胡兴旺,易朵,王成,赵晓蒙,周畅..重叠延伸PCR法构建小鼠Sumf1基因的定点突变真核表达载体[J].生命科学研究,2009,13(5):430-436,7.基金项目
国家自然科学基金资助项目(30500258) （30500258）
湖南省教育厅科研资助项目(09B059) （09B059）
