The Wallabies will travel to Kingsholm on Tuesday, November 3, four days before they take on England at Twickenham and three days after they face New Zealand in Tokyo.
The fixture will be the first time an Australian national side has played at the famous West Country Stadium in more than a century.
The Australians, who were nicknamed the rabbits by British media on their 1908/09 tour, were 6-5 winners against the Cherry and Whites last time out.
That tour also saw the side coin the moniker the 'Wallabies', with the tourists voting for that name ahead of other contenders such as Waratahs, Wallaroos and Kookaburras.
And with this November's tour the first for 25 years in which the Wallabies will play all four of the Home Unions, the added trip to Kingsholm provides even more excitement surrounding an already hectic schedule.
"The history surrounding this tour is compelling and we're delighted a match against Gloucester will deepen the links to some great Wallabies teams of the past," said Australian Rugby Union managing director and CEO John O'Neill.
"This will be the first Grand Slam attempt since the 1984 squad became the first Australians to triumph against the four Home Unions of England, Ireland, Wales and Scotland on the same tour."
And while O'Neill was quick to highlight the historical importance of the fixture, Wallaby head coach Robbie Deans made it clear that the mid-week game will help his young-look squad build up for what he expects to be four demanding international Test matches in London, Dublin, Edinburgh and Cardiff.
"We want to provide players with the opportunity to wear the national jersey and build competition for places in the Test side," said Deans.
"The match against Gloucester will fulfill that objective and give our players an early taste of British conditions against one of the leading clubs in Europe."
For English fans, the game offers a chance to see the new generation of Wallabies first-hand, two years ahead of the 2011 World Cup and four years out from the 2013 Lions tour Down Under.
Wallabies tour schedule
October 31: vs All Blacks - Olympic Stadium, Tokyo, 8.30am GMT
November 3: vs Gloucester - Kingsholm, 7.45pm
November 7: vs England - Twickenham, 2.30pm
November 15: vs Ireland - Croke Park, 3pm
November 21: vs Scotland - Murrayfield, 5.15pm
November 28: vs Wales - Millennium Stadium, 5.15pm