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Endo-β-glucanase Ⅱ (EG Ⅱ) gene cDNA was isolated from the fungus Humicola insolens H31-3 by RT-PCR.It was cloned into the expression vector pGAPZαA.The resultant recombinant plasmid was introduced into Pichia pastoris GS115 by electroporation after being lin-earized by BspHI digestion.The recombinant Pichia pas-toris strain was obtained and SDS-PAGE showed that the molecular weight of the expression protein was about 55 kD.The cultivation condition and the characteristics of the recombinant EG Ⅱ were also explored.