LZ-8基因的克隆及其在毕赤酵母(Pichia pastoris)中的表达

Cloning and expression in Pichia pastoris of LZ-8 gene from the Gamoderma lucidum

用 PCR 从灵芝(Gamoderma lucidum)基因组扩增免疫蛋白 LZ-8基因,将其构建到酵母表达质粒 pPICZaA 中,采用电激转化法转化毕赤酵母,获得转化子。对其 Sourthern 杂交分析,结果表明 LZ-8基因整合到毕赤酵母基因组。对转化酵母作发酵培养,SDS—聚炳烯酰胺凝胶电泳检测到 LZ-8蛋白的表达。

LZ-8 gene was isolated and cloned from the genome DNA of the Gamoderma lucidum。LZ-8 gene fragments were successfully amplified by PCR reactions.The gene is inserted into expression vector pPICZaA,and give rise to plasmid pPLZ8-X.Then Pichia yeasts were transformed by electrophoration with pPLZ8-X,and were selected on medium containing Zeocin.Southern blot shows that LZ-8 gene has integrated into the yeast ge- nome.By incubating the transformant of Pichia yeast,LZ-8 protein was purified from the culture supernatant and detected by SDS-PAGE.