The 2000 Southern Miss Golden Eagles football team represented the University of Southern Mississippi in the 2000 NCAA Division I-A football season.
The Golden Eagles were led by head coach Jeff Bower and played their home games at M. M. Roberts Stadium.
For the second time in three years, the Golden Eagles earned a preseason top 25 ranking, coming in at No.
The Golden Eagles win streak ended that week against eventual conference champions Louisville, and they would go onto lose two more games to finish the regular season with a 7–4 record, 4–3 in Conference USA play.
Despite the late season struggles, the Golden Eagles earned a trip to the Mobile Alabama Bowl, where they defeated 13th-ranked TCU, 28–21.