Israeli authorities on Thursday closed the Kerem Shalom crossing for the fourth day in a row, an official said Maan reported.
Raed Fattouh, a border official, told Ma'an that Israeli authorities informed the Palestinian side that the crossing would be closed for security reasons.
Israel sealed its border with the Gaza Strip on Monday after militants fired a rocket into the Negev desert on Sunday evening.
Police spokesman Micky Rosenfeld told AFP the rocket landed in an uninhabited sector of the Negev desert.
Israeli Defense Minister Moshe Yaalon warned on Wednesday that his country would respond to any attacks on its territory and not allow its people to come under fire.
MP Jamal al-Khodri condemned the border closure as an attempt to tighten the siege and collapse the Palestinian economy.
Rights groups have criticized Israeli restrictions on civilian travel and goods transfer in response to rocket fire as collective punishment.
After Israel closed the Gaza border in March following rocket fire, Gisha, an Israeli organization that campaigns for Palestinian freedom of movement, wrote to the defense minister.
"In the last month, there appears to be a new policy toward the Gaza Strip, in which Israel is openly restricting civilian movement to and from Gaza, not because of a concrete security necessity, but rather as a punitive step taken against the civilian population in direct response to fire by combatants," wrote Gisha director Sari Bashi.