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白纹伊蚊唾液三磷酸腺苷二磷酸酶在毕赤酵母中的分泌表达 被引量：1

Secretory Expression of Salivary ATP Diphosphohydrolase(Apyrase)from Aedes albopictus in Pichia pastoris
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摘要 采用RT-PCR技术克隆编码白纹伊蚊(Aedes albopictus)唾液三磷酸腺苷二磷酸酶(apyrase)的成熟肽cDNA序列,并克隆至毕赤酵母(Pichia pastoris)组成型分泌表达载体pGAPZα-A的α-factor信号肽序列的下游,构建pGAPZα-A-apyrase重组分泌表达载体。表达载体经BlnⅠ线性化处理后电击转化毕赤酵母GS115感受态细胞,转化子经Zeocin抗性筛选和菌落PCR,成功构建了pGAPZα-A-apyrase/GS115工程菌。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDSPAGE)结果显示,pGAPZα-A-apyrase/GS115的工程菌分泌表达了相对分子质量(M_r)约为60000的重组apyrase蛋白。 The gene-coding mature apyrase protein from Aedes cdbopictus was amplified by RT-PCR and cloned in frame with the α-factor secretion signal peptide into Pichia pastoris secreting expression vector pGAPZα-A resulting in the pGAPZα-A-apyrase. After being linearized by Bln I restriction enzyme, the recombinant pGAPZα-A-apyrase was trans-formed into Pichia pastoris GSll5 by electroporation. Recombinant strains pGAPZα-A-apyrase/GSll5 were screened on YPDS plates containing Zeocin and identified by PCR. The recombinant protein of apyrase （Mr 60 000） has been expressed in the supernatant of Pichia pastoris.
作者 吴松泉 王光丽 雷永良 张凯波
机构地区 丽水学院医学院 丽水市疾病控制中心
出处 《中国寄生虫学与寄生虫病杂志》 CAS CSCD 北大核心 2013年第1期73-75,共3页 Chinese Journal of Parasitology and Parasitic Diseases
关键词 白纹伊蚊 ATP-二磷酸酶 毕赤酵母 表达 Aedes albopictus Apyrase Pichia pastoris Secreted expression
分类号 R384.113 [医药卫生—医学寄生虫学]
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参考文献12

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二级参考文献51

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中国寄生虫学与寄生虫病杂志
2013年 第1期

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