父杜鄴曾任涼州(今甘肅省張家川回族自治縣)刺史。王莽末年,盜賊蜂起,杜林和弟杜成避難於河西(今甘肅武威),隗囂聽說杜林有志節,打算讓他做持書平。杜林不肯,又勸他歸順東漢,隗囂不聽。杜林以弟杜成去世為由,請求攜柩回鄉安葬。隗囂大怒,派刺客楊賢去殺他。楊賢見到杜林用鹿車推亡弟的靈柩,感念其友孝,嘆說「當今之世,誰能行義?我雖小人,何忍殺義士?」,遂不忍殺之。建武六年(30年)歷官侍御史,官至大司空。杜林精研孔子之禮,曾於河西得漆書《古文尚書經》一卷,隨着帶在身邊,後來賈逵爲之作《訓》,馬融作《傳》,鄭玄《註解》,古文尚書開始流傳。
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