Justice Secretary Michael Gove is to run to be the next Conservative Party leader and UK prime minister.
Mr Gove, a prominent figure in the Brexit campaign, had been expected to support Boris Johnson's candidacy.
He said he was standing because he had come "to the conclusion that Boris cannot provide the leadership or build the team for the task ahead".
Home Secretary Theresa May, who backed staying in the EU, has also entered the contest. Nominations close at noon.
The winner of the contest is set to be announced on 9 September.
The leadership battle has been sparked by David Cameron's to step down as prime minister after losing the EU referendum, which saw the country vote by 52% to 48% to leave the EU.
Mr Johnson, the former mayor of London, is widely expected to enter the race with a speech later, and to place Brexit at the heart of his "optimistic vision" for the country.
He is expected to say the greater self-determination that leaving the EU will bring is an opportunity to "believe in ourselves and the values of our country".
Energy minister Andrea Leadsom, who also campaigned to leave the EU, has also thrown her hat in to the ring, tweeting: "Let's make the most of the Brexit opportunities!"
She joins former Defence Secretary Liam Fox and Work and Pensions Secretary Stephen Crabb among the runners and riders.
Health Secretary Jeremy Hunt has pulled out of a leadership bid and given his backing to Mrs May, as has Education Secretary Nicky Morgan who had been considering running herself.
'Unity and change'
Mr Gove's announcement that he will challenge the leadership was unexpected, as the justice secretary had been expected to throw his weight behind fellow leading Leave campaigner Mr Johnson for Conservative leader.
Explaining his decision, he said: "I have repeatedly said that I do not want to be prime minister. That has always been my view. But events since last Thursday have weighed heavily with me.
"I respect and admire all the candidates running for the leadership. In particular, I wanted to help build a team behind Boris Johnson so that a politician who argued for leaving the European Union could lead us to a better future.
"But I have come, reluctantly, to the conclusion that Boris cannot provide the leadership or build the team for the task ahead."
Who's in the running?
Home Secretary Theresa May: The 59-year-old has overtaken Boris Johnson as the bookies' favourite to win the contest. She's held the Home Office brief - often something of a poisoned chalice - since 2010, and is a former Tory party chairman. She says she can offer the "strong leadership" and unity the UK needs, and promised a "positive vision" for the country's future. She backed staying in the EU.
Former London mayor Boris Johnson: The 52-year-old journalist-turned-Conservative politician is one of the most familiar faces in British politics, his unconventional political style and unique brand of charisma making him a household name. He spent eight years in City Hall before re-entering Parliament last year - and was the most prominent figure in the winning Brexit campaign.
Justice Secretary Michael Gove: The 48-year-old former newspaper columnist was a key figure in the party's modernisation that led to its return to power in 2010. He was a reforming, if controversial, education secretary between 2010 and 2014, and now holds the Ministry of Justice brief. He was a leading player in the Brexit campaign - which put a strain on his close friendship with David Cameron. He has pitched himself as the candidate that can provide "unity and change."
Work and Pensions Secretary Stephen Crabb: The 43-year-old was promoted to the cabinet in 2014 as Welsh secretary, and boosted his profile earlier this year when he took over as work and pensions secretary. A rising star of the Tory party he has promised to unite the party and country following the referendum result and provide stability. Raised on a council estate by a single mother, he has a back story to which many Tory MPs are attracted. Backed Remain.
Energy minister Andrea Leadsom: The 53-year-old former banker and fund manager was one of the stars of the Leave campaign. A former district councillor, she became MP for South Northamptonshire in 2010 and - after serving as a junior Treasury minister and as a member of the Treasury select committee - she was made a junior minister in the energy and climate change department in May last year.
Former cabinet minister Liam Fox: It's second time around for the 54-year-old ex-defence secretary and GP, who came a close third in the 2005 leadership contest. His cabinet career was cut short in 2011 when he resigned following a lobbying row. A Brexit campaigner, and on the right of the party, he has said whoever becomes PM must accept "the instruction" of the British people and not "try to backslide" over EU membership.
Setting out his pitch for the leadership, the cabinet minister - who was formerly education secretary - said: "I want there to be an open and positive debate about the path the country will now take.
"Whatever the verdict of that debate I will respect it. In the next few days I will lay out my plan for the United Kingdom which I hope can provide unity and change."
The UK faced "huge challenges" but also "huge opportunities", he said, and added: "If we are to make the most of the opportunities ahead we need a bold break with the past."
Conservative MP David Davis said Mr Gove's decision must have been taken "very late", as Mr Gove's assistant had asked him on Wednesday night to attend Mr Johnson's campaign launch on Thursday.
"So clearly there are some very late decisions going on in all of this," told BBC Radio 5 Live.
Meanwhile, justice minister and Leave campaigner Dominic Raab has switched his support from Mr Johnson to Mr Gove, saying he has the "vision and technical mastery" for the task ahead.
"He's going to be the underdog but he will be fighting for the underdog," Mr Raab said.
'Bold, positive vision'
Launching her leadership bid in central London, Mrs May - one of the longest-serving home secretaries in history - said the UK needed "strong proven leadership to steer us through this period of economic and political uncertainty and to negotiate the best possible terms as we leave the European Union".
Although she backed Remain she maintained a low profile during the campaign and, in her speech, insisted she would not back away from last week's vote. "Brexit is Brexit."
But she said Article 50 - the formal mechanism for leaving the EU - should not be triggered until the UK had agreed its negotiating strategy - probably before the end of year.
She set out plans for a new government department to oversee the UK's withdrawal from the EU - and said it would be headed by a pro-Brexit cabinet-level minister.
Mrs May also said that, as prime minister she would drop her call for Britain to withdraw from the European Convention on Human Rights, saying she did not expect there to be a parliamentary majority for it.
Mrs May - the daughter of a vicar - also pitched herself as the candidate that could "unite our party and our country".
And she pledged "a bold, new positive vision for the future of our country" that "works not for a privileged few, but for every one of us".
Mrs May acknowledged she was "not a showy politician", adding: "I just get on with the job in front of me," - suggesting that was what the country wanted.
In a swipe at potential leadership rival Mr Johnson, joking: "Boris negotiated in Europe. I seem to remember last time he did a deal with the Germans, he came back with three nearly-new water cannon."
The then London mayor purchased the riot control water cannon following the riots in the capital city in 2011 - but Mrs May eventually withheld permission for their use.
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