The Montenegro international came close to completing a move to the Turin club in the summer, but the deal eventually collapsed as Juve failed to reach a compromise with Fiorentina.
The Italian champions are keen to bolster their strikeforce, and were also linked with Liverpool forward Luis Suarez and Manchester United striker Robin Van Persie before he joined the Old Trafford club.
And, despite their interest in Jovetic over the summer, they are now no longer keen on the Montenegrin.
"We spoke with Fiorentina president [Mario] Cognigni and sporting director [Daniele] Prade about Jovetic," said Juventus director Giuseppe Marotta.
"However, we are no longer interested in signing him now."
The termination of interest from Juventus opens the door for Manchester City, who were linked with the striker for much of the summer.
Roberto Mancini is thought to be a big fan of the forward, and Manchester City are reportedly preparing a €30m bid for the striker in January.
The Premier League champions signed Matija Nastasic from Fiorentina in the summer, and Manchester City could go back to secure the transfer of Jovetic once the transfer window reopens.
Jovetic has started the season in impressive form, bagging four goals in his first six league games.