The Norfolk Southern Railroad was founded on New Year's Day 1883, with its original road running from Norfolk, Virginia, to Elizabeth City, North Carolina. Its predecessor, the Elizabeth City & Norfolk Railroad, chartered in 1870, had completed tracks south to Edenton on the Albemarle Sound in 1881. This was a continuation of a commercial connection between these points dating back to the Dismal Swamp Canal efforts of the late 1700s. In 1891 the road was reorganized as the Norfolk & Southern Railroad, but after 1900 consolidations brought back the name Norfolk Southern and the system was extended to New Bern, Raleigh, and Charlotte.

In 1907 work commenced on a five-mile trestle to cross the sound, which by January 1910 cost more than $1 million. The railroad lacked sufficient funds to adequately maintain the trestle, which perhaps led to its collapse in 1957-an accident that dropped a train into the sound and killed two crew members. But the trestle opened the way to connect with points in Piedmont North Carolina. Several of the state's smaller lines became part of the Norfolk Southern, including the Winston-Salem Southbound Railway, Raleigh & Western Railway (originally the Egypt Railway), Raleigh & Cape Fear Railway, North Carolina Midland Railroad, and Charlotte, Columbia, and Augusta Railroad.

The line eventually ran from Norfolk to both Charlotte and Greensboro via Raleigh. In 1961 the company offices were moved from Norfolk to Raleigh.

The Norfolk Southern was noted for transporting agricultural and lumber products, and it aided in industrial development, especially in the rural counties of eastern North Carolina. At times the railroad offered passenger service, but after 1948 it was completely a freight road.

Never strong financially, the company went into receivership on several occasions. On January 1, 1974 the Norfolk Southern united with the Southern Railway System but continued to operate under its own name. At the time of the merger, the Norfolk Southern consisted of nearly 1,000 cars and engines and 624 miles of track. In 1982, when Southern Railway merged with the Norfolk & Western Railroad, the railroad became a part of the massive Norfolk Southern Corporation.