用显微注射方法分析牙鲆肌肉发育调节基因MyoD, Myf5的启动子序列

克隆了牙鲆(Paralichthys olivaceus)肌肉发育调节基因MyoD和Myf5的启动子序列,分别为608 bp和1 073 bp。对序列进行分析发现MyoD和Myf5的启动子含有参与肌肉发育调控的基因的结合位点。将MyoD和Myf5的启动子连接到GFP载体上,构建了重组质粒MyoDP-GFP和Myf5P-GFP,并注射到一细胞或两细胞的斑马鱼胚胎中,对这两个基因的启动子进行了瞬时表达研究。结果表明,牙鲆MyoD和Myf5的启动子可以驱动绿色荧光蛋白特异地在斑马鱼肌肉纤维中表达,因此牙鲆608 bpMyoD和1073 bpMyf5的启动子包含着可以驱动MyoD和Myf5正常表达的必需元件,它们是肌肉特异的,并且可以跨物种行使其功能。

Characterization of promoters from flounder(Paralichthys olivaceus) muscle-regulatory genes, MyoD and Myf5

The promoter sequences of flounder muscle regulatory gene,MyoD and Myf5,are cloned as 608 bp and 1 073 bp respectively.Many muscle regulatory gene binding sites are found in the promoters of MyoD and Myf5.The promoter sequences were ligated to GFP vector and constructed as MyoDP-GFP and Myf5P-GFP.The result constructs were microinjected into one-cell or two-cell stage zebrafish embryos to make a transient expression analysis.The results showed that the 608 bp MyoD promoter and the 1 073 bp Myf5 promoter can drive GFP specifically expressed in muscle fibers in zebrafish.It can be seen that the two promoters contain muscle specific regulatory elements,and they can function across species.