Gateway cloning is compatible with protein secretion from Pichia pastoris

被引：8

作者：

Esposito, D

Gillette, WK

Miller, DA

Taylor, TE

Frank, PH

Hu, RQ

Bekisz, J

Hernandez, J

Cregg, JM

Zoon, KC

Hartley, JL

机构：

[1] SAIC Frederick Inc, Natl Canc Inst, Protein Express Lab, Frederick, MD 21702 USA

[2] NIAID, NIH, Bethesda, MD 20892 USA

[3] Keck Grad Inst Appl Life Sci, Claremont, CA 91711 USA

来源：

PROTEIN EXPRESSION AND PURIFICATION | 2005年 / 40卷 / 02期

关键词：

Pichia pastoris; gateway cloning secretion; interferon;

D O I：

10.1016/j.pep.2004.12.006

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Secretion of a recombinant protein from the yeast Pichia pastoris requires the presence of a signal peptide at the amino terminus. Maintaining the full amino acid sequence of the signal peptide is thought to be important for proper signal processing and protein secretion. We show that at least for one protein, a synthetic human interferon, the presence of a Gateway recombination site within the signal peptide is fully compatible with high levels of protein secretion. The amino termini of the secreted interferon proteins cloned with Gateway and cloned with restriction enzymes and ligase are identical, and the proteins were highly active in biological assays. Compatibility with Gateway cloning simplifies construction of plasmids directing secretion of recombinant proteins from P. pastoris. (c) 2005 Elsevier Inc. All rights reserved.

引用

页码：424 / 428

页数：5

共 10 条

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