GroEL, a viral binding protein encoding by groEL gene from endosymbionts in Bemisia tabaci, is an important factor in transmission of plant viruses. In this study, 502 bp fragment of groEL was amplified by polymerase chain reaction (PCR) and then ligated to the plant expression vector pBIN19. The recombination vector pBIN19::groEL was transformed into Agrobacterium tumefaciens strain LBA4404 by electroporation. Transgenic Nicotiana benthamiana plants were acquired by Agrobacterium-mediated approach. PCR and RT-PCR were performed to analyze groEL’s transformation and expression in transgenic plants, respectively. The results indicated that groEL gene was not only integrated into chromosome DNA of transgenic N. benthamiana, but also transcribed in transgenic plants successfully.