摘要
目的:构建雌激素受体a(ERa)的真核表达载体并在293T细胞内进行表达鉴定。方法:以乳腺文库为模板,PCR扩增ERa基因序列,回收片段,BamHI、XhoI双酶切片段及pcDNA3.0载体,将其连接并转化到DH5a感受态细胞中,酶切鉴定阳性克隆,转染293T细胞进行westernblot实验,检测pcDNA3.0-ERa载体的表达。结果:获得了雌激素受体a的真核表达载体pcDNA3.0-ERa。结论:本研究成功构建了雌激素受体a的真核表达载体pcDNA3.0-Era,为研究雌激素及其受体在心血管系统中的作用提供了基础。
Objective： To construct estrogen receptor a （ERa） eukaryotic expression vector pCDNA3.0-ERa and identified the vector in 293T cells. Methods： ERa gene was amplified from breast library by PCR. And the product was digested by restriction enzymes BamHI and XhoI. The product was ligated with eukaryotic expression vector pcDNA3.0 digested by BamHI and XhoI. Then the expression vector was ransfected to DH5a cells and positive vectors digested by BamHI and XhoI was chosen. pcDNA3.0-ERa was transfected to 293T cells and the expression was identified by western blot. Results： A Estrogen receptor eukaryotic expression vector pcDNA3. 0-ERa was obtained. Conclusion： An estrogen receptor eukaryotic expression vector pcDNA3.0-ERa was successfully constructed.
出处
《现代生物医学进展》
CAS
2010年第19期3620-3622,共3页
Progress in Modern Biomedicine
