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    The HepG2 cells transfected with pCAT3-Basic was used as negative control. The activity of CAT inHepG2 cells transfected was detected by an enzyme-linked immunosorbent assay (ELISA) kit after 48 hours, which reflected the trans-regulating function of bicyclol on cyclin B2p gene promoter.
    以该质粒转染肝癌细胞系HepG2细胞,用酶联免疫吸附法(ELISA)检测氯霉素乙酰转移酶(CAT)的表达活性并与双环醇共刺激的HepG2细胞,用ELISA法检测CAT的表达活性。
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  p genes
The objective was to relate the response of the HSP70 and P53 genes to the cessation and the recovery of cardiac muscle cell functions when submitted to ischemia-reperfusion.
      
Comparative study of the expression Rb and p53 genes in human colorectal cancers, color carcinoma cell lines and synchronized hu
      
Comparative study of the expression of Rb and p53 genes in human colorectal cancers, colon carcinoma cell lines and synchronized
      
Concurrent abnormal expression of ERBB-2, EGFR, and p53 genes and clinical disease progression of breast carcinoma
      
These p53 genes were individually mutated at amino acid residues 143, 175, 248 and 273.
      
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Objective To investigate the trans-regulating effect of bicyclol on cyclin B2 gene promoter.Methods Polymerase chain reaction (PCR) technique was employed to amplify the sequence of cyclin B2 promoter by using HepG2 genomic DNA as template, and the amplified product was subcloned into pCAT3-Basic at Kpn I and Bgl 11 sites, and the resulted plasmid was designated pCAT3-cyclin B2p. pCAT3-cyclin B2p was transfected into the hepatoblastoma cell line HepG2 by FuGENE 6 transfection re agents, then...

Objective To investigate the trans-regulating effect of bicyclol on cyclin B2 gene promoter.Methods Polymerase chain reaction (PCR) technique was employed to amplify the sequence of cyclin B2 promoter by using HepG2 genomic DNA as template, and the amplified product was subcloned into pCAT3-Basic at Kpn I and Bgl 11 sites, and the resulted plasmid was designated pCAT3-cyclin B2p. pCAT3-cyclin B2p was transfected into the hepatoblastoma cell line HepG2 by FuGENE 6 transfection re agents, then stimulated with bicyclol. The HepG2 cells transfected with pCAT3-Basic was used as negative control. The activity of CAT inHepG2 cells transfected was detected by an enzyme-linked immunosorbent assay (ELISA) kit after 48 hours, which reflected the trans-regulating function of bicyclol on cyclin B2p gene promoter. Results The report vector pCAT3-cyclin B2p has been constructed and had been confirmed by restriction enzyme digestion and sequencing. The expression of CAT in HepG2 cells co-transfected with pCAT3-cyclin B2p and bicyclol was 2.4 times as higher as that of pCAT3-Basic, and 0.35 as higher as that of pCAT3-cyclin B2p.Conclusion Bicyclol candown-regulate cyclin B2 promoter.

目的探讨双环醇对细胞周期素(cyclin)B2启动子转录活性的调节作用。方法根据文献报道的结果确定细胞周期素B2的启动子DNA序列区域,以聚合酶链反应(PCR)扩增细胞周期素启动子(B2p),克隆至真核报告载体pCAT3Basic中,构建pCAT3cyclinB2p报告载体;以该质粒转染肝癌细胞系HepG2细胞,用酶联免疫吸附法(ELISA)检测氯霉素乙酰转移酶(CAT)的表达活性并与双环醇共刺激的HepG2细胞,用ELISA法检测CAT的表达活性。结果成功获得细胞周期素B2启动子的止确克隆。pCAT3cyclinB2p和双环醇(106Mol/L)瞬时转染的HepG2细胞的CAT表达活性是pCAT3Basic空载体的2.4倍,pCAT3cyclinB2p的0.35倍。结论细胞周期素B2启动子有顺式激活下游基因的活性,双环醇具有对细胞周期素B2基因有下调作用。

 
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