Tory revolt was led by Thatcherite 'class of 2010'

More than half of the dissident MPs who joined the biggest rebellion on Europe ever suffered by a Tory prime minister were only elected last year.

The scale of the insurgency among the "class of 2010", which is demonstrating greater independence of spirit than any previous group of new MPs from any party, will alarm Downing Street.

David Cameron insisted yesterday there was "no bad blood, no rancour, no bitterness" on his part following Monday night's revolt, in which 81 Tory MPs defied a three-line whip. But the ramifications from the result continued yesterday, as Downing Street confirmed that two ministerial aides, Adam Holloway and Stewart Jackson, who backed a motion calling for a referendum on Europe, had been "removed" – but not "sacked" – from their posts.

David Nuttall, the MP for marginal Bury North, has emerged as the rebel-in-chief. He has an unorthodox, working-class background among Conservatives – he was educated at a comprehensive school, left school at the age of 18 and later obtained a law degree by correspondence.

His hard slog, fighting as a councillor in the Tory-unfriendly territory of south Yorkshire, has given him the doggedness and independence that led to him tabling the motion that caused his leader so much grief.

Other high-profile rebel newcomers include Zac Goldsmith (Richmond Park), Mark Reckless (Rochester and Strood), Priti Patel (Witham), a former spokeswoman for William Hague when he was party leader, and Sarah Wollaston, the GP chosen as the Tory candidate in Totnes following an open primary.

Many of the new Conservative MPs had their political awakening during the Thatcher years, with the result that they share her instincts on Europe, as well as on tax and reining in the public sector.

They also arrived after a wave of public fury over the expenses scandal swept out hundreds of MPs at the last election – and are less likely to share their predecessors' deference to appeals for party loyalty.

Almost 50 of Monday night's rebels were first elected in 2010. They joined forces with traditional members of the Tory awkward squad, such as Douglas Carswell, Philip Davies, Philip Hollobone and Peter Bone, and longer-standing right-wingers instinctively hostile to Mr Cameron's leadership, most notably David Davis, Christopher Chope and Edward Leigh.

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