The first works will involve the excavation of the track alignment for the new line through the Monktonhall area and the excavation of the site of the new Shawfair station.
An earthworks programme, which will create the line of route and base for the railway track, will now begin to roll-out along the route throughout summer and autumn.
A start to main works follows a six-month period of advance preparations. This included a great deal of mining remediation work in the area of the old Monktonhall colliery.
Scottish Transport Minister Keith Brown said: “Our delivery of the Borders Railway is bringing communities along the route back into the rail network for the first time in over 40 years.
“The start of the construction works is a hugely exciting milestone and will allow those communities to witness the scheme coming to life.”
The Borders Railway is due to enter service in 2015 and will re-establish a passenger rail link for the first time in over 40 years from Edinburgh through Midlothian to Tweedbank.
The line will include 30 miles of new railway with three existing stations and seven new ones – at Shawfair, Eskbank, Newtongrange, Gorebridge, Stow, Galashiels and Tweedbank.