This is the first immigration-related vote in either chamber of Congress this year and a measure of the daunting challenge facing supporters of a sweeping overhaul of existing law on the subject.
The party-line vote of 224-201 was aimed at blocking implementation of President Barack Obama's 2012 election-year order called the DREAM Act to stop deportations of many individuals who could benefit from it.
Democrats on the House floor reacted with boos when the provision was added to a routine spending bill for the Department of Homeland Security.
It nevertheless stood as a stark warning from conservatives who dominate the ranks of the Republican House majority about attempts in the Senate to grant a chance at citizenship to an estimated 11 million immigrants residing in the country illegally.
And the White House reacted sharply, saying the House-passed measure would affect "Dreamers" who are "productive members of society who were brought here as young children, grew up in our communities, and became American in every way but on paper."
Rep. Steve King, a Republican, said in a statement that the vote prohibits the administration "from implementing executive amnesty" without congressional action.
My amendment blocks many of the provisions that are mirrored in the Senate's 'Gang of Eight' bill. If this position holds, no amnesty will reach the President's desk," he said.
The vote took place as Senate leaders set Friday for the opening of debate on White House-backed legislation that would create a chance at citizenship for those in the country unlawfully, at the same time it takes steps to assure the borders are secure against future illegal immigration.
The measure was drafted by a bipartisan group of eight senators, then approved by the Senate Judiciary Committee last month on a vote of 13-8.
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