关键词: Notch信号, NTM, 人胰腺癌, 真核表达载体

Abstract: Objective To construct the eukaryotic expression vector of human Notch1 transmembrane domain(NTM), pcDNA3-NTM, and to examine its expression in vitro. Methods Total RNA was extracted from the BxPC3 cells and reverse transcription-polymerase chain reaction(RT-PCR) was performed to obtain the cDNA fragment encoding Notch1 transmembrane domain, which was inserted into the pcDNA3 vector in-frame with the flag sequence. And the new construct was confirmed by restriction enzyme digestion and DNA sequencing. HeLa cells were transfected with the pcDNA3-NTM vector and pcDNA3 vector, respectively. The expression of NTM was detected by Western blotting. Results The eukaryotic expression vector pcDNA3-NTM was constructed, and significant increase of NTM expression was detected in the HeLa cells 48 hours after transfection. Conclusion The eukaryotic expression vector pcDNA3-NTM was constructed successfully.

Key words: notch signaling, notch1 transmembrane domain, human pancreatic cancer, eukaryotic expression vector