重复序列竞争反式调节因子模型的构建和初步研究

The construction and research of the model of repeated sequences competing trans-regulation factors

目的构建不同拷贝数的重复串联序列表达质粒,研究串联重复序列对表达质粒中GFP报告基因表达的影响作用。方法将不同拷贝数的2F2R、AC和AT串联重复序列插入pEGFP-Alu14或pEGFP-C1中,构建表达载体,瞬时转染HeLa细胞,提取RNA做Northern印记检测。结果在4个拷贝数范围以内,2F2R随拷贝数增加活化基因作用增强,再增加拷贝数其活化基因作用反而下降;装有重复序列的质粒随转染剂量上升表达先上升后下降。结论重复序列可能通过竞争反式调节因子来调节基因表达。

Objective We constructed expression plasmids of repeated tandem sequences of different copy number and researched the influence of GFP reporter gene expression by repeated tandem sequences. Methods We constructed Expression vectors by inserting repeated tandem sequences of 2F2R, AC and AT of different copy number into pEGFP-Alu14 or pEGFP-C1, transfected with HeLa cells and extracted RNA to commit Northern blot Results With the scope of 4 copies, the effect of activating gene of 2F2R increased following the increase of copy number, but following copy number was increased again, the effect of activating gene of 2F2R decreased instead. the expression of plasmids inserted with repeated sequences increased firstly and decreased later. Conclusion The repeated sequences may regulate gene expression by competing trans-regulation factors.