【摘要】 目的:构建Neuritin的毕赤酵母分泌表达系统,为进一步探讨Neuritin的生物学功能及作用机制奠定基础。方法和结果:在已克隆Neuritin cDNA的基础上,PCR扩增出Neuritin ORF与载体pPIC9k重组,然后利用电击转化将重组质粒pPIC9k-Neuritin导入GS115酵母菌株。筛选出阳性克隆经PCR及测序正确后,用甲醇诱导Neuritin分泌表达;表达产物经SDS-PAGE及Western-blot鉴定。结论:成功构建Neuritin的毕赤酵母分泌表达系统。更多还原

【Abstract】 Objective: Construct Pichia pastoris secretory system of Neuritin,set a base for making a further research on function and mechanism of Neuritin.Methods and results:We amplified Neuritin open reading frame from cloned Neuritin cDNA by PCR and inserted it into pPIC9k vector,then transform pPIC9k-Neuritin recombinant into Pichia pastoris GS115.After identifying positive clone by PCR analysis and sequencing,the recombinant was induced by methanol to express Neuritin.The expressed products were analysised by SDS-PAGE and Western-blot.Conclusion:We successfully constructed Pichia pastoris secretory system of Neuritin.更多还原