CMV启动子与艾美耳球虫基因侧翼序列对黄色荧光蛋白表达调控作用的研究

Regulation of Yellow Fluorescent Protein Expression by CMV Promoter and Genetic Flanking Sequences of Eimeria

探索CMV启动子与艾美耳球虫基因侧翼序列对串联型黄色荧光蛋白(YFP)表达的调控作用,将含有串联型YFP、CMV启动子、柔嫩艾美耳球虫组蛋白4(H istone4)上游启动序列和肌动蛋白(Actin)下游调控序列的重组载体pCMV-YFP-YFP、pH4-YFP-YFP-ACT电转染Vero细胞和柔嫩艾美耳球虫子孢子,观察荧光情况。结果证明,CMV启动子不能驱动YFP在柔嫩艾美耳球虫中正常表达,而其自身基因的侧翼序列能够启动和调控串联型YFP在柔嫩艾美耳球虫中的正常表达。

To explore the regulation of tandem repeated yellow fluoresent protein（YFP） expression in Vero ceils .and sporozoites of Eimeria, the recombinant vectors pCMV - YFP - YFP which contains tandem repeated YFP and CMV promoter, and pH4 - YFP - YFP - ACT which contains tandem repeated YFP too, upstream flanking sequence of Histone4 （H4） and downstream regulatory sequence of Actin both derived from E. tenella genomic DNA, were transfected into Vero cells and sporozoites of E. tenella by electroporation. Yellow fluorescence within cells and parasites were observed by virtue of fluorescence microscope, Which indicated that genetic flanking sequences of E. tenella could promote functional expression of tandem YFP gene in E. tenella , on contrary, CMV promoter could not.