The Senate voted 51-49 in favor of a procedural motion on the bill, which would spend $60 billion on transportation and infrastructure. However, 60 votes were required to break a GOP filibuster on the bill.
It was the third time in recent weeks Republicans were able to block measures that had been components of the American Jobs Act, initially proposed by the administration as a comprehensive, $447 billion package intended to boost hiring and give money to states to hire teachers and public safety workers.
After the vote, the president said it was "more clear than ever that Republicans in Washington are out of touch with Americans from all ends of the political spectrum."
"The American people deserve to know why their Republican representatives in Washington refuse to put some of the workers hit hardest by the economic downturn back on the job rebuilding America," Obama said -- repeating what has become a constant White House theme.
"It's time for Republicans in Congress to put country ahead of party and listen to the people they were elected to serve," he said in a statement. "It's time for them to do their job and focus on Americans' jobs. And until they do, I will continue to do everything in my power to move this country forward."
Many Republicans said they favored transportation and infrastructure projects but objected to a provision that would fund the projects with a new tax on income over $1 million, The Hill.com reported.