糙皮侧耳漆酶基因的克隆及在粟酒裂殖酵母中的表达

Cloning and Expression of a Pleurotus ostreatus Laccase Gene in Schizosaccharomyces pombe

利用RT-PCR技术,从糙皮侧耳(Pleurotus ostreatus)总RNA中克隆漆酶基因(Lac),再通过NheⅠ酶切位点与粟酒裂殖酵母(Schizosaccharomyces pombe)表达载体pESP-2连接,构建酵母真核表达载体,利用电转化方法转化粟酒裂殖酵母SP-Q01细胞,并在SP-Q01中进行表达分析,最后用愈创木酚法测定表达产物的酶学活性。漆酶基因在构建的表达载体重组SP-Q01细胞中得到表达,酶学活性达到89.37 U/L。

A laccase gene from Pleurotus ostreatus was cloned using RT-PCR, inserted into the expressing vector pESP-2 of Schizosaccharomyces pornbe to generate a recombinant plasmid pESP-2-Lac, and the plasmid was transformed into S. pornbe SP-QO1 by electroporation. Laccase levels of 89.37 U/L were recorded in culture supernatants of the transformant S. pornbe-Lac