Glyphosate is a highly efficient, broad-spectrum nonspecific herbicide that inhibits the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS)-mediated pathway of shikimic acid.  The screening of glyphosate-resistant EPSPS gene is a major means for the development of new genetically modified glyphosate-resistant transgenic crop.  Currently, the main commercialized glyphosate-resistant soybean contains glyphosate-resistant gene CP4-EPSPS.  In this study, a G10-EPSPS gene was reported providing glyphosate resistance in Zhongdou 32.  Here, G10-EPSPS gene was introduced into soybeans through Agrobacterium-mediated soybean cotyledon node.  PCR, Southern blotting, semi-quantitative RT-PCR, qRT-PCR, and Western blotting were used, and the results revealed that G10-EPSPS had been integrated into the soybean genome and could be expressed steadily at both mRNA and protein levels.  In addition, glyphosate resistance analysis showed that the growth of transgenic soybean had not been affected by concentrations of 900 and 2 700 g a.e. ha^–1 of glyphosate.  All the results indicated that G10-EPSPS could provide high glyphosate resistance in soybeans and be applied in production of glyphosate-resistant soybean.