Israel's military said that two Israeli soldiers have been killed by a driver from Jordan, bringing in humanitarian aid for Gaza.
Israel's Magen David Adom ambulance service said the two Israelis, who initially were not identified, succumbed to their wounds while the attacker was shot dead by security forces.
The attack took place at the King Hussein Bridge, also known as the Allenby Bridge, at the border crossing from Jordan into the occupied West Bank.
The Israel Defense Forces (IDF) said that it recommended the government halt humanitarian aid arriving from Jordan pending an investigation into "the inspection procedures for the Jordanian drivers."
Jordan's Foreign Ministry condemned the attack in a statement and said an investigation would be launched, saying the shootings were a threat to its humanitarian role in Gaza.
4 Israeli soldiers killed in Gaza explosion
Israel's military reported the deaths of four soldiers in southern Gaza, the first deaths reported since the launch of an offensive to seize Gaza City.
Israeli media reported that the four were killed in the early hours of the morning in the southern Gazan city of Rafah.
The deaths come as Israeli warplanes pounded targets in parts of the city.
The AFP news agency cited journalists and witnesses as saying they saw a steady stream of Palestinians heading south on foot, in vehicles and on donkey carts with piles of their belongings.
The UN estimated at the end of August that around 1 million people were living in Gaza City and its surroundings. Israel says 350,000 of them have fled.