摘要
目的通过转录因子Sp1基因克隆以及真核表达载体的构建，为探讨转录因子Sp1在牙釉质发育的分子调控奠定基础。方法根据引物设计原则设计Sp1基因的PCR引物，以从小鼠成釉细胞中提取的总RNA逆转录所得eDNA为模板，进行PCR扩增，用PCR法扩增得出含有EcoRⅠ和XbaⅠ的Sp1目的基因连接到pcDNA3．1／myc-HisA真核表达载体上。结果经过PCR引物扩增得到2343bp基因片段，将获得的重组质粒pcDNA3．1／myc-HisA-Sp1双酶切分析鉴定，测序结果与GenBank登录基因完全一致。结论成功实现了Sp1基因克隆及表达载体的构建，为以后研究转录因子Sp1在牙釉质发育的分子调控方面奠定基础。
Objective To study the significance of Spl in developing dental enamel,through the transcription factor Spl gene cloning and constnwt eukaryotic expression vector cDNA3.1/myc-HisA of transcription factor Spl ,and lay a foundation. Methods According to the PCR primer design principles,the eDNA coding sequence of Spl containing the digestion site of EcoR Ⅰ and Xba Ⅰ was amplified by RT-PCR frum ameloblasts and then was inserted into pcDNA3.1/myc-HisA vector. Results The amplified gene fragment was 2343bp. Re- striction enzyme digestion and sequencing showed that pcDNA3, l/myc-HisA-Spl was constructed correctly. And it was confirmed by sequen- cing which was consistent with samples in GenBank. Conclusion The Spl gene cloning and eukaryotic expression vector Spl is constructed successfully,and to lay the foundation for future studies of transcription factor Spl in the molecular regulation of enamel development.
出处
《潍坊医学院学报》
2012年第3期189-191,I0003,共4页
Acta Academiae Medicinae Weifang
基金
国家自然科学基金资助课题（课题编号：30973327）,山东省自然科学基金资助课题（课题编号：ZR2010HM076）
