利用跨越内含子的ＰＣＲ技术，从三角酵母（Ｔｒｉｇｏｎｏｐｓｉｓｖａｒｉａｂｉｌｉｓ）变种ＦＡ１１０中扩增得到Ｄ氨基酸氧化酶基因（ｄａａｏ），并通过ＴＡ克隆的方法将其克隆至ｐＧＥＭＴ载体。序列测定结果表明，所得ｄａａｏ基因的５′端内含子已被删除，基因总长度为１０７１ｂｐ，它与Ｔｒｉｇｏｎｏｐｓｉｓｖａｒｉａｂｉｌｉｓ的Ｄ氨基酸氧化酶同源性达９８３％，与Ｆｕｓａｒｉｕｍｓｏｌａｎｉ和Ｒｈｏｄｏｔｏｒｕｌａｇｒａｃｉｌｉｓ的同源性分别是３８９％和３０８％。为提高表达水平，又将此基因转移至高表达载体ｐＥＴ２８ｂ上，在大肠杆菌ＢＬ２１（ＤＥ３）中进行诱导表达。经ＩＰＴＧ诱导，目的蛋白的产生量可占菌体总蛋白量的４６％，分子量约为３８ｋＤ。Ｄ氨基酸氧化酶的活力可达８０２ｕ／Ｌ。 The Dao amino acid oxidase gene (daao) was amplified from the Trigonopsisvariabilis variety FA1-10 by using intron-crossing PCR technique and cloned into pGEM-T vector by TA cloning method. Sequence analysis showed that the 5 ’end of intron of daao gene was deleted. The total length of the gene was 1071bp. Its homology with Damino acid oxidase of Trigonopsisvariabilis was 983%, which was close to that of Fusarium solani and Rhodotoru  lagracilis The homologies were 389% and 308% respectively. In order to increase the expression level, this gene was transferred to the high expression vector pET28b and induced in E. coli BL21 (DE3). After induced by IPTG, the amount of the target protein can account for 46% of the total amount of the bacterial protein and the molecular weight is about 38 kD. D amino acid oxidase activity up to 802u / L.