一个新葡萄糖淀粉酶基因的克隆与表达

Cloning and Expression of a New Glucoamylase Gene

摘要:

根据已报道的米根霉葡萄糖淀粉酶基因序列，通过PCR方法，从天然少根根霉的总DNA中克隆到含有四个内含子的葡萄糖淀粉酶基因。通过设计引物并采取重叠PCR方法删除内含子，获得了新的少根根霉葡萄糖淀粉酶(Rhizopus arrhizu glucoamylase,RaGA)cDNA序列(Accession number：DQ903853)。该基因在毕赤酵母中成功表达，表达产物具有较高的葡萄糖淀粉酶活性。

Abstract:

According to the reported gene sequence of Rhizopus oryzae glucoamylases, the glucoamylase gene containing four introns was cloned from the total DNA of the natural Rhizopus arrhizu. Specific primers were designed to delete introns by overlapping PCR and a new cDNA sequence of Rhizopus arrhizu glucoamylase was obtained. The accession number in gene bank is DQ903853. This gene is successfully expressed in the Picha pastoris, producing a new protein with a high activity of glucoamylase.