The variety of conductive fibers has been constantly enriched in recent years, and it has made rapid development in the fields of electronic textiles, intelligent wearable, and medical care. However, the environmental damage caused by the use of large quantities of synthetic fibers cannot be ignored, and there is little research on conductive fibers in the field of bamboo, a green and sustainable material. In this work, we used the alkaline sodium sulfite method to remove lignin from bamboo, prepared a conductive bamboo fiber bundle by coating a copper film on single bamboo fiber bundles using DC magnetron sputtering, and analyzed its structure and physical properties under different process parameters, finding the most suitable preparation condition that combines cost and performance. The results of the scanning electron microscope show that the coverage of copper film can be improved by increasing the sputtering power and prolonging the sputtering time. The resistivity of the conductive bamboo fiber bundle decreased with the increase of the sputtering power and sputtering time, up to 0.22 Ω·mm; at the same time, the tensile strength of the conductive bamboo fiber bundle continuously decreased to 375.6 MPa. According to the X-ray diffraction results, Cu in the copper film on the surface of the conductive bamboo fiber bundle shows the preferred orientation of (111) the crystal plane, indicating that the prepared Cu film has high crystallinity and good film quality. X-ray photoelectron spectroscopy results show that Cu in the copper film exists in the form of Cu0 and Cu2+, and most are Cu0. Overall, the development of the conductive bamboo fiber bundle provides a research basis for the development of conductive fibers in a natural renewable direction.