【摘要】 利用跨越内含子的PCR技术,从三角酵母(Trigonopsisvariabilis)中扩增得到D-氨基酸氧化酶基因(DAAO),通过TA克隆的方法将其克隆至pBS-T载体,并转化大肠杆菌TOP10。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。之后将该基因与酵母表达载体pPIC3.5K连接,转化巴斯德毕赤酵母(Pichiapastoris)。更多还原

【Abstract】 The D-amino acid oxidase gene(DAAO) was cloned from the total DNA of Trigonopsis vairabilis by intron deletion PCR amplification The PCR fragment of DAAO was ligated to a T-vector for DNA sequencing.It was shown that the PCR fragment was about 1071 bp without an intron domain at 5’-terminal of DAAO.Then the fragment was inserted in pPIC3.5K and transformed into Pichia pastoris.更多还原