The long-awaited update supersedes a list from early 2025, and comes less than a month after President Donald Trump met China's Xi Jinping on a visit to Beijing, where the two leaders maintained a delicate trade war truce.
China's foreign ministry said on Tuesday that the list was discriminatory and "unreasonably suppressed" Chinese companies, urging the US to "correct its mistaken practices".
"China will take necessary measures to firmly safeguard the legitimate rights and interests of Chinese enterprises," ministry spokesman Lin Jian told reporters at a briefing on Tuesday.
The list now includes a broad swathe of China's top technology firms key to advancing Beijing's military and industrial prowess, reflecting Washington's security concerns amid intense geopolitical competition between the countries.
In February, when Trump's trip to China had been pending, the Pentagon briefly posted an updated list, known as the 1260H or CMC list, but then quickly withdrew it with little explanation.
The new version released on Monday mirrors the withdrawn February list with the exception of the inclusion of China's top memory chipmakers CXMT and YMTC, two companies that had been removed from the short-lived February index to the ire of Washington's China hawks.
Other companies added include biotech firm WuXi AppTec , AI-driven robotics company RoboSense Technology Co Ltd and Unitree, a leading Chinese maker of humanoid and quadruped robots.
On June 1, US AI chipmaker Nvidia said it plans to work with Unitree to build robots for researchers.
BYD, the world's largest EV seller, said in a statement to Reuters that it firmly opposed being labelled a military company and would use all "feasible administrative and legal means" to safeguard its rights and interests, adding that the decision harmed "its development achievements in the United States".
Alibaba said there was "no basis" for its inclusion on the list.
"Alibaba is not a Chinese military company nor part of any military-civil fusion strategy. We will take all available legal action against attempts to misrepresent our company," the e-commerce and tech conglomerate said.
WuXi AppTec responded that its inclusion on the list was "incorrect", and said in a statement that it would "take immediate actions to challenge and correct this erroneous designation."
Search engine and AI giant Baidu "categorically" rejected its inclusion on the list, and in a statement said: "The suggestion that Baidu is a military company is entirely baseless. We will not hesitate to use all options available to us to have the company removed from the list."
