LZ-8基因克隆及其在毕赤酵母中的诱导表达

【目的】从灵芝中克隆LZ-8基因并在毕赤酵母表达系统中诱导表达LZ-8蛋白。【方法】采用聚合酶链反应(PCR)法从赤灵芝的基因组DNA中扩增LZ-8基因序列,采用电激转化法将pPLZ8-X重组质粒转化到毕赤酵母中,PCR Southernblot法检测酵母是否转化LZ-8基因,十二烷基磺酸钠—聚丙烯酰胺凝胶电泳(SDS-PAGE)并银染法观察蛋白在酵母中的差异表达。【结果】PCR Southern blot结果显示LZ-8基因已转化到酵母中;SDS-PAGE胶银染结果提示LZ-8蛋白在酵母中被诱导表达。【结论】免疫调节蛋白基因LZ-8在毕赤酵母表达系统获得了诱导表达。

LZ-8 Gene Cloning from Ganoderma lucidum and Its Inducible Expression in Pichia pastoris

Objective To clone immune modulator LZ-8 gene from Ganoderma lucidum and to induce the expression of LZ-8 protein in Pichia pastoris.Methods LZ-8 gene was amplified from the genomic DNA of Ganoderma lucidum by polymerase chain reaction(PCR),and pPLZ8-X recombinant vector harboring LZ-8 gene was transformed into Pichia pastoris by electroporation.PCR Southern blot method was used to detect the presence of LZ-8 gene in Pichia pastoris,sodium dodecyl sulphate-polyacrylamide gel electrophoresis(SDS-PAGE) and subsequent silver staining were applied to display the differential protein expression in Pichia pastoris.Results The result of PCR Southern blot showed the presence of LZ-8 gene in Pichia pastoris,and silver staining results indicated that LZ-8 gene expression was induced in Pichia pastoris.Conclusion LZ-8 gene cloned from Ganoderma lucidum has been expressed in Pichia pastoris which belongs to a potent eukaryotic expression system.