PHOⅠ信号肽在毕赤酵母中引导分泌表达天然N-端rBPTI

doi:国家高技术研究发展计划（863计划）资助课

PHOⅠ信号肽在毕赤酵母中引导分泌表达天然N-端rBPTI

杨莉莉¹，贺巾超¹，马杰¹，范伟全¹，王建秋¹，魏传宇¹，颜浩为²，颜炜群^1*

1. 吉林大学再生医学科学研究所生物化学教研室，吉林长春130021;2.吉林大学药学院生物工程系，吉林长春 130021

收稿日期:2005-04-25 修回日期:1900-01-01 出版日期:2005-11-28 发布日期:2005-11-28
通讯作者: 颜炜群

Expression and secretion of natural rBPTI with PHOⅠ signal peptide in Pichia pastoris

YANG Li-li¹, HE Jin-chao¹, MA Jie¹, FAN Wei-quan¹, WANG Jian-qiu¹, WEI Chuan-yu¹, YAN Hao-wei²,YAN Wei-qun^1*

1. Department of Biochemistry, Institute of Frontier Medical Sciences, Jilin University, Changchun 130021, China;2. Department of Bioengineering, School of Pharmacy, Jilin University, Changchun 130021, China

Received:2005-04-25 Revised:1900-01-01 Online:2005-11-28 Published:2005-11-28
Contact: YAN Wei-qun

摘要/Abstract

摘要： 目的：在毕赤酵母(Pichia pastoris)中分泌表达天然N-端rBPTI。方法：将PHOⅠ/bpti基因克隆到真核表达载体pPICZα，电转化毕赤酵母菌株X-33。结果：构建了含PHOⅠ/bpti基因的表达质粒,并在 X-33中分泌表达rBPTI。用胰蛋白酶抑制实验筛选出表达rBPTI的工程菌。表达上清经阳离子交换层析分离纯化为单一组份，SDS-PAGE显示相对分子质量为 6 500。结论：PHOⅠ信号肽在毕赤酵母中成功地诱导分泌表达了具有正确N-端氨基酸序列的rBPTI。

关键词: 酸性磷酸酶信号肽, 毕赤酵母

Abstract: Objective To express and secrete the natural N-terminal rBPTI with PHOⅠ signal peptide in Pichia pastoris. Methods PHOⅠ/bpti genes were inserted into the eukaryotic expression plasmid. The recombinant plasmid was transformed into the Pichia pastoris (X-33) via electroporation. Results The expression plasmid was constructed to contain correct sequence for PHOⅠ/bpti genes. The rBPTI was expressed and secreted in X-33. An activity strain was selected with trypsin inhibitor experiment .The expression supernatant was purified with cation exchange chromatography to single peak and SDS-PAGE indicated that the relative molecular mass was 6 500. Conclusion rBPTI with natural N-terminal sequence is successfully expressed in Pichia pastoris with signal peptide of PHOⅠ.

Key words: acid phosphatase signal peptide, Pichia pastoris

中图分类号:

Q516

杨莉莉，贺巾超，马杰，范伟全，王建秋，魏传宇，颜浩为，颜炜群. PHOⅠ信号肽在毕赤酵母中引导分泌表达天然N-端rBPTI[J]. J4, 2005, 31(6): 830-832.

YANG Li-li, HE Jin-chao, MA Jie, FAN Wei-quan, WANG Jian-qiu, WEI Chuan-yu, YAN Hao-wei,YAN Wei-qun. Expression and secretion of natural rBPTI with PHOⅠ signal peptide in Pichia pastoris[J]. J4, 2005, 31(6): 830-832.

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