菊糖果糖转移酶基因在毕赤酵母中的分泌表达  

Secretory expression of inulin fructotransferase in Pichia pastoris

作　　者：战荣荣[1] 沐万孟[1] 江波[1] 周榴明[1] 

机构地区：[1]江南大学食品科学与技术国家重点实验室,江苏无锡214122

出　　处：《食品工业科技》2013年第1期171-174,共4页Science and Technology of Food Industry

摘　　要：将PCR获得的不包含信号肽序列的菊糖果糖转移酶基因(ift)与毕赤酵母分泌表达载体pPIC9K进行连接,构建重组载体pPIC9K-IFTase。重组载体经线性化后电转入毕赤酵母GS115感受态细胞,利用G418抗性梯度筛选及PCR鉴定,获得一株高拷贝菊糖果糖转移酶重组毕赤酵母菌株。该重组菌株经甲醇诱导,能够分泌具有活性的菊糖果糖转移酶,且60h时酶活为10.3U/mL。结果表明,菊糖果糖转移酶可以实现在毕赤酵母的分泌表达。The ift gene without signal peptide sequence was cloned through PCR and ligated with the expression vector pPIC9K of P.Pichia,resulting in the recombinant plasmid pPIC9K-IFTase.The recombinant plasmid was then linearized and transformed into P.pastoris GS115,and a hight copy recombinant strain of yeast was acquired through concentration gradient resistance screening of G418,and PCR identification method.Using methanol induction for 60h,the recombinant strain P.pastoris secreated IFTase having an activity of 10.3U/mL.This finding indicated that Eukaryotic secretory expression of the IFTase could be performed in P.pastoris.

关 键 词：双果糖酐Ⅲ 毕赤酵母 分泌表达 真核表达 

分 类 号：Q786[生物学—分子生物学]