Journal of Capital Medical University ›› 2009, Vol. 30 ›› Issue (4): 500-503.doi: 10.3785/j.issn.1006-7795.2009.04.020
• 基础研究 • Previous Articles Next Articles
ZHANG Kai-ju, YU He-fen, ZHANG Yu-xiang
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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
CLC Number:
R 34
ZHANG Kai-ju;YU He-fen;ZHANG Yu-xiang. Construction and Identification of the Eukaryotic Expression Vector of Human Notch1 Transmembrane Domain[J]. Journal of Capital Medical University, 2009, 30(4): 500-503.
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URL: https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/10.3785/j.issn.1006-7795.2009.04.020
https://journal03.magtech.org.cn/Jweb_sdykdxxb/EN/Y2009/V30/I4/500