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Neuritin在毕赤酵母中的分泌表达与鉴定
Secretary Expression and Identification of Neuritin in Pichia Pastoris
【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。
【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.
【Key words】 Neuritin; pPIC9k; Pichia pastoris; gene expression;
- 【文献出处】 农垦医学 ,Journal of Nongken Medicine , 编辑部邮箱 ,2006年05期
- 【分类号】Q786
- 【被引频次】1
- 【下载频次】108