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- W2397675911 abstract "Objective To analyze the transcription activation and possible regulation mechanism of human X-box binding protein 1 (XBP1) gene 5′upstream DNA sequence in different cell lines. Methods Six kinds of XBP1 promoter deletion mutants were cloned into pGEM-Teasy vector, which included XBP1 gene 5′ upstream -1039 to 66 bp,-859 to 66 bp,-623 to 66 bp,-351 to 66 bp,-227 to 66 bp,-227 to -45 bp respectively. Every deletion mutant sequence was cut from Teasy-XBP1p by Kpn Ⅰ and Xho Ⅰ, and subcloned into pCAT3-Basic to produce a set of constructs termed as p1-XBP1p, p2-XBP1p, p3-XBP1p, p4-XBP1p, p5-XBP1p, p6-XBP1p, respectively. The transcription activity of each construct was detected after transiently transfecting K562, HepG2,NIH-3T3 and L02 cell with FuGENE 6 transfection reagent. Cells transfected by pCAT3-Basic or pCAT3-Promoter were used as negative and positive controls. The activity of chloramphenicol acetyltransferase(CAT), which reflects the transcription activation of the XBP1 gene promoter, was detected by ELISA after 48 hours of transfection. Results The reporter vectors of six kinds ofXBP1 promoter deletion mutants were successfully constructed, as confirmed by restriction enzyme digestion and sequencing. The activities of p4-XBP1p and p5-XBP1p were higher than the other deletion mutants in K562 and HepG2. And the activity of p5-XBP1p was the highest in HepG2. There was no activity detected from any transfected NIH-3T3. Conclusion The XBP1 gene promoter can transactivate its downstream gene to transcription. The core sequence ofXBP1 promoter was implied between -227 bp and 66 bp. This sequence was connected with the transcriptional activity ofXBP1 promoter closely. Its transcription activity varies with different cell lines.XBP1 promoter might drive gene expression with cell-type specificity.Key words: X-box binding protein 1; promoter; chloramphenicol acetyltransferase reporter gene; transcriptional regulation" @default.
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- W2397675911 date "2006-02-01" @default.
- W2397675911 modified "2023-09-23" @default.
- W2397675911 title "An analysis on transcriptional regulation activity of human XBP1 gene 5' upstream DNA sequences." @default.
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