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John V of Anhalt-Zerbst (Dessau, 4 September 1504 – Zerbst, 4 February 1551), was a German prince of the House of Ascania and ruler of the principality of Anhalt-Dessau. From 1544, he assumed rule of the re-created principality of Anhalt-Zerbst. John was the second (but eldest surviving) son of Ernest I, Prince of Anhalt-Dessau, by his wife Margarete, daughter of Henry I, Duke of Münsterberg-Oels, and granddaughter of George of Poděbrady, King of Bohemia.