The Applications of GFP as a Reporter Gene in Rice Genetic Transformation

In this study,a plasmid pJPM5 containing modified GFP gene which encode the green fluorescent protein (GFP) was constructed.Microprojectile bombardment was used to separately introduce two plasmids pJPM5 and pSBG700 into rice TNG67 calli.The transgenic plants were confirmed by Southern blot.Most of the transgenic plants contain 1 to 8 copies of transgene.GFP gene expression was measured in the extracts from rice leaf of two month old transgenic plants.And the quantification of gfp expression is carried by the SLM-8000 Fluorescence Analyzer. Most of the transgenic plants have showed significant GFP signal. Even though the rice plant has auto fluorescence, the expression of GFP is high enough to give out much higher fluorescence signal compared with auto fluorescence of rice plants.The GFP was also observed in transformed calli under microscope.The transient expression of GFP helped to demonstrate that the bombardment shots of 900psi 1350 psi make the particles carrying plasmids reach more cells than the bombardment shots of 2X 900psi or 2X 1350psi. Our results showed that this modified GFP gene can be used as a reporter gene for rice genetic transformation and even for other cereal crops such as wheat and corn etc.The result also professes that the MAR sequence can enhance the expression of GFP gene significantly(data was shown in another paper).