安徽医科大学学报2012，Vol.47Issue(12)：1377-1380,4.
真核载体pcDNA3.1/PSD95-PDZ2的构建、表达及功能鉴定
Construction, detection of expression and function of eukaryotic vector pcDNA3.1/PSD95-PDZ2
摘要
Abstract
Objective To construct the eukaryotic vector pcDNA3. 1/PSD95-PDZ2, and detect its expression and function in PC12 cells. Methods The 309 bp fragment of PSD95-PDZ2 was amplified by RT-PCR from the cDNA of neurons. The fragment and the vector pcDNA3. 1 were digested with restriction enzymes EcoR I and BamH I, and the digested productions were connected by T4 DNA ligase at 16℃ , and then the eukaryotic expression vector of pcDNA3. 1/PSD95-PDZ2 was constructed. The plasmid was identified by the double digestion with restriction enzymes EcoR I and BamH I and DNA sequencing. After the analysis, pcDNA3. 1/PSD95-PDZ2 was transfected into PC12 cells by Lipofectamine? 2000, and the expression and function of PSD95-PDZ2 were detected by IP. Results The PSD95-PDZ2 fragment was contained in the positive recombination by identification of restriction enzymes, and the sequence was the same on the GenBank. The 11 ku-sized protein was detected by IP, which proved that PSD95-PDZ2 could express and combine with nNOS in the infected PC12 cells. Conclusion The eukaryotic vector of pcDNA3. 1/PSD95-PDZ2 has been constructed successfully, and the fragment of PSD95-PDZ2 has been expressed and combined with nNOS in PC12 cells,which paves the way for further studies on the functions of PSD95-PDZ2.关键词
PSD95-PDZ2/真核表达载体/构建/表达Key words
PSD95-PDZ2/ eukaryotic expression vector/ construct/ expression分类
生物科学引用本文复制引用
朱明媚..真核载体pcDNA3.1/PSD95-PDZ2的构建、表达及功能鉴定[J].安徽医科大学学报,2012,47(12):1377-1380,4.基金项目
国家自然科学基金(编号:30971021) （编号:30971021）
