转铁蛋白（TF）是一种具有重要生理功能的糖蛋白，对它的开发利用具有重要的经济和社会效益。为探讨高效表达人转铁蛋白（hTF）的可行性及可能的调控机理,本文克隆得到了兔转铁蛋白启动子（RP promoter），并构建了以pcDNA3.1(-)为骨架的由RP启动hTF表达的重组载体，转染大鼠肝脏细胞株BRL-3A和小鼠乳腺上皮细胞株HC-11细胞分析了hTF表达情况，并进一步筛选得到了两个稳定表达hTF的细胞克隆。在细胞培养液中加入不同剂量的5-杂氮胞苷（5-aza-dC）,分析甲基化修饰对基因表达的影响。结果显示,兔转铁蛋白启动子能成功启动hTF的表达；而且5-aza-dC能进一步提高了hTF表达水平，且提高的程度和5-aza-dC的剂量成正相关，但是不同克隆hTF表达升高程度不一样。结果说明hTF的表达受到甲基化修饰影响。
To achieve high level expression of human transferring (hTF) and investigate the possible mechanism of the hTF expression, rabbit transferrin promoter (RP promoter) was cloned, and a hTF recombinant expression vector (δpcDNA3.1(-)-RP-hTF), was constructed in which hTF DNA was driven by RP promoter. The transient transfection experiments achieved a high hTF expression in BRL-3A and HC-11 cell lines, and two stably integrated cell clones were also screened out. Various concentrations of 5-aza-dC were added to the cell culture medium for the investigation of the effect on hTF expression. Real-time RT-PCR analysis showed that the RP promoter directed an efficient hTF expression, and 5-aza-dC increased hTF expression in a proportional manner. However, the extent of the increase in hTF expression differed between individual clones, suggesting that TF expression may relate to gene methylation.
潘树标,颜景斌,任兆瑞. 人转铁蛋白重组载体的构建及其表达调控的研究[J]. 中国生物工程杂志, 2008, 28(专刊): 50-55.
Yan Jing-Bin . Construction of Human transferrin (hTF)Recombinant Vector and the Regulation of Its Expression. China Biotechnology, 2008, 28(专刊): 50-55.
https://manu60.magtech.com.cn/biotech/CN/ 或 https://manu60.magtech.com.cn/biotech/CN/Y2008/V28/I专刊/50
Cited
