The acceleration of the anthropogenic activity has increased the atmospheric carbon concentration, which causes changes in regional climate. The Gross Primary Production (GPP) is an important variable in the global carbon cycle studies, since it defines the atmospheric carbon extraction rate from terrestrial ecosystems. The objective of this study was to estimate the GPP of the Amazon-Cerrado Transitional Forest by the Vegetation Photosynthesis Model (VPM) using local meteorological data and remote sensing data from MODIS and Landsat 5 TM reflectance from 2005 to 2008. The GPP was estimated using Normalized Difference Vegetation Index (NDVI) and Enhanced Vegetation Index (EVI) calculated by MODIS and Landsat 5 TM images. The GPP estimates were compared with measurements in a flux tower by eddy covariance. The GPP measured in the tower was consistent with higher values during the wet season and there was a trend to increase from 2005 to 2008. The GPP estimated by VPM showed the same increasing trend observed in measured GPP and had high correlation and Willmott's coefficient and low error metrics in comparison to measured GPP. These results indicated high potential of the Landsat 5 TM images to estimate the GPP of Amazon-Cerrado Transitional Forest by VPM.