Police say a semi-tractor trailer overturned near mile marker 90 just after 11:30 Sunday morning.
Traffic has been backed up for several miles.
Motorists are encouraged to seek an alternate route.
There is no word on when the lane will be reopened.
The truck driver was not hurt in the accident.
Crews have also set up a lane closure on southbound I-65 from mile marker 26 to mile marker 19 until 7:00 Monday morning for contruction purposes.