With the development of modern molecular biology, clone and expresson technique is improved. For the study the mechanism of GDNF promoting the proliferation of glioma cells, many genes need to knock down or over-expression. So we need an effective protein expression system. In this study we use Gateway Technology, which is easy for us to construct a expression vector.. Promoter is a very important element for a protein expression, because some promoters maybe not promote the expression of the protein in glioma cells, some promoters maybe too strong, which will kill the cell after transformation. So we systematicly compare nine promoters which we have. After comparison, we found the CMV, SV40, PGK promoters are all work well in glioma cells, and the pgk is the best one. BetaActin dosen’t work well, which will cause glioma cells to die.