Spain's rider Alberto Contador won this year's Giro but will not defend his title
Lance Armstrong's first Giro d'Italia will begin with a team time trial in Venice and finish with an individual time trial in Rome.
The route for the race, which celebrates its centenary this year, was unveiled on Saturday in Venice.
It takes place from 9-31 May and includes five mountain stages and a time trial on the Ligurian coast.
The race usually finishes in Milan but for the centennial event it will reach its climax in the capital.
Armstrong is returning to cycling next year after retiring in 2005 and the seven-time Tour de France winner is planning to race both the Giro and Tour for the first time in his career.
"I am very excited to come to Italy to participate in my first Giro and I look forward to a good result," he said.
"I began my professional career living near Lake Como, but I never competed in the Giro.
"This is a special year for many reasons. Obviously, there is the hundred year anniversary of the race, but it's also a special route from Venice to Rome.
There will be a lot of dynamics in an exciting first week while the second week is highlighted by the long time trial of 62 km and the last week has some tricky stages.
"Rome is a city that I never experienced until last year, when I visited the city as a tourist and I fell immediately in love with it."
Milan will host the finish of the ninth stage and Mount Vesuvius in Naples will be the site of a mountain stage two days before the finish in Rome.
The route will also pass through several points connected with the Giro's history, and cover many of Italy's biggest cities - Venice, Milan, Florence, Bologna and Naples.
It will venture into the French Alps during the 10th stage, which will begin and finish in Italy.
This year's winner Alberto Contador announced in October that he would not be taking part in next year's event but former winners Ivan Basso, Danilo Di Luca, Damiano Cunego and Gilberto Simoni are set to race again.
This year's Tour de France winner Carlos Sastre has also entered along with British sprinter Mark Cavendish.
Giro d'Italia 2009 stages
Stage 1 - Saturday 9 May, Lido di Venezia, 20.5km Team time trial
Stage 2 - Sunday 10 May, Jesolo - Trieste, 156km
Stage 3 - Monday 11 May, Grado - Valdobbiadene, 200km
Stage 4 - Tuesday 12 May, Padova - San Martino di Castrozza, 165km
Stage 5 - Wednesday 13 May, San Martino Di Castrozza - Alpe di Siusi, 125km
Stage 6 - Thursday 14 May, Bressanone / Brixen - Mayrhofen (Aut), 242km
Stage 7 - Friday 15 May, Innsbruck (Aut) - Chiavenna, 244km
Stage 8 - Saturday 16 May, Morbegno - Bergamo, 208km
Stage 9 - Sunday 17 May, Milano Show 100, 155km
Rest day - Monday 18 May
Stage 10 - Tuesday 19 May, Cuneo - Pinerolo, 250km
Stage 11 - Wednesday 20 May, Torino - Arenzano (Genova), 206km
Stage 12 - Thursday 21 May, Sestri Levante - Riomaggiore, 61.7km individual time trial
Stage 13 - Friday 22 May, Lido di Camaiore - Firenze, 150km
Stage 14 - Saturday 23 May, Campi Bisenzio - Bologna (San Luca), 174km
Stage 15 - Sunday 24 May, Forli - Faenza, 159km
Rest day - Monday 25 May
Stage 16 - Tuesday 26 May, Pergola - Monte Petrano, 237km
Stage 17 - Wednesday 27 May, Chieti - Blockhaus, 79km
Stage 18 - Thursday 28 May, Sulmona - Benevento, 181km
Stage 19 - Friday 29 May, Avellino - Vesuvio, 164km
Stage 20 - Saturday 30 May, Napoli - Anagni, 203km
Stage 21 - Sunday 31 May, Roma, 15.3km individual time trial