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哺乳动物非经典分泌信号肽介导重组EGFP蛋白向毕赤酵母细胞壁转运

覃玉凤1 陈瑶生1 刘志国1 张英1 刘海龙1 何祖勇1

(1.中山大学生命科学学院有害生物控制与资源利用国家重点实验室, 广东广州 510006)

【摘要】为探索哺乳动物非经典分泌信号肽在毕赤酵母表达系统中引导重组蛋白分泌的作用,本研究将一段来源于小鼠同源异型框蛋白(En2)的分泌信号序列(SS)融合至EGFP蛋白的N端,在毕赤酵母中表达。实验结果显示SS信号肽能通过一种不同于经典的内质网-高尔基体分泌通路的方式将EGFP蛋白分泌至细胞膜表面,与α交配因子前导肽相比,显著降低了细胞的内质网压力。本研究提示哺乳动物非经典分泌信号肽可作为递送重组蛋白至酵母膜表面的一项工具。

【关键词】 内质网压力;非经典分泌;毕赤酵母;qPCR;

【DOI】

【基金资助】 国家转基因生物新品种培育重大专项(No.2016ZX08006003-006); 广东省自然科学基金(No.2016A030313310)资助;

Delivery of recombinant enhanced green fluorescent protein to Pichia pastoris cell wall directed by a mammalian nonclassical secretion signal peptide

Qin Yufeng 1 Chen Yaosheng 1 Liu Zhiguo1 Zhang Ying 1 Liu Hailong 1 He Zuyong 1

(1.State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou, Guangdong, China 510006)

【Abstract】A mammalian nonclassical secretion sequence derived from mouse Engrailed 2 homeoprotein (En2) was used to direct the secretion of the enhanced green fluorescent protein from Pichia pastoris. This signal peptide conferred the transport of enhanced green fluorescent protein into periplasm through an endoplasmic reticulum-golgi independent pathway, without inducing severe unfolded protein response as compared with Saccharomyces cerevisae α-factor preprosequence. This study implies that this mammalian nonclassical signal peptide could be developed as a useful tool for delivering cargoes to the cell surface of yeast.

【Keywords】 ER stress; nonclassical secretion; Pichia pastoris; qPCR;

【DOI】

【Funds】 National Transgenic Major Program (No. 2016ZX08006003-006); Natural Science Foundation of Guangdong Province (No. 2016A030313310);

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ISSN:1000-3061

CN: 11-1998/Q

Vol 32, No. 10, Pages 1455-1464

October 2016

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Abstract

  • 1 Materials and methods
  • 2 Results and analysis
  • 3 Discussion
  • References