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使用新型G418抗性载体直接筛选毕赤酵母表达菌株。

Direct selection of Pichia pastoris expression strains using new G418 resistance vectors.

作者信息

Lin-Cereghino Joan, Hashimoto Matthew D, Moy Allison, Castelo James, Orazem Claire C, Kuo Peter, Xiong See, Gandhi Vishal, Hatae Christopher T, Chan Alex, Lin-Cereghino Geoff P

机构信息

Department of Biological Sciences, University of the Pacific, Stockton, CA 95211-0197, USA.

出版信息

Yeast. 2008 Apr;25(4):293-9. doi: 10.1002/yea.1587.

DOI:10.1002/yea.1587
PMID:18327886
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2504081/
Abstract

The methylotrophic yeast, Pichia pastoris, is widely used as a host organism for the expression of heterologous proteins. Currently, the Zeocin and blasticidin resistance genes are the only dominant selectable markers that can be used for primary selection of transformants. In this report we describe new expression vectors that can be used to select directly for P. pastoris transformants using G418 resistance conferred by a modified Tn903kan(r) gene. Compared to other dominant markers, this system is more economical and offers a higher transformation efficiency, due to the small sizes of the cloning vectors, pKAN B and pKANalpha B (GenBank Accession Nos EU285585 and EU285586, respectively). Additionally, multicopy transformants can be generated using these new vectors.

摘要

甲基营养型酵母毕赤酵母被广泛用作表达异源蛋白的宿主生物。目前，博来霉素和杀稻瘟菌素抗性基因是仅有的可用于转化体初步筛选的显性选择标记。在本报告中，我们描述了新的表达载体，这些载体可利用经修饰的Tn903kan(r)基因赋予的G418抗性直接筛选毕赤酵母转化体。与其他显性标记相比，由于克隆载体pKAN B和pKANalpha B（GenBank登录号分别为EU285585和EU285586）尺寸较小，该系统更经济且转化效率更高。此外，使用这些新载体可产生多拷贝转化体。

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