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毕赤酵母菌株的高通量筛选与选择

High-Throughput Screening and Selection of Pichia pastoris Strains.

作者信息

Weis Roland

机构信息

VALIDOGEN (formerly VTU Technology) GmbH, Raaba-Grambach, Austria.

出版信息

Methods Mol Biol. 2019;1923:169-185. doi: 10.1007/978-1-4939-9024-5_7.

Abstract

Clone screening procedures for Pichia pastoris expression strain comparison rely on the availability of a cultivation environment that ensures equal growth and production capabilities for all assessed transformants. As clonal variation in such experiments is caused by diverging numbers and possibly also genomic locations of integrated (linearized) expression constructs, the productivity assessment of a larger number of strains is mandatory for selecting a set of strains for follow-up bioreactor cultivations in order to define the best-producing clone. Microscale cultivation provides the means to reliably compare growth and productivity of a large number of transformants and by that narrows down the amount of selected strains for scaling up.

摘要

用于毕赤酵母表达菌株比较的克隆筛选程序依赖于一种培养环境,该环境要确保所有评估的转化体具有同等的生长和生产能力。由于此类实验中的克隆变异是由整合(线性化)表达构建体数量的差异以及可能的基因组位置差异引起的,因此为了选择一组菌株用于后续的生物反应器培养以确定最佳生产克隆,必须对大量菌株进行生产力评估。微尺度培养提供了可靠比较大量转化体生长和生产力的方法,从而减少用于扩大规模的选定菌株数量。

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