The funding, from Silicon Valley venture capital firm Kleiner Perkins Caufield & Byers and existing Twitter investors, underscores the high hopes investors have for social networking sites.
Twitter, which has more than 175m users, says it will use the money to further expand its operations.
The moves come just two months after the four-year-old company handed the job of chief executive to Dick Costolo, the architect of its new advertising efforts, a sign that making money is a priority for the service.
Costolo said he plans to have hundreds of advertisers using Twitter’s ad system by the end of the year.
Twitter had raised $160m in four earlier funding rounds, from investors that included Spark Capital, Institutional Venture Partners and mutual fund company T Rowe Price.
However, the site still has a long way to go to catch up with Facebook, which has more than 500m users and a valuation of more than $45bn.