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模式绿藻莱茵衣藻的遗传转化

Genetic transformation of the model green alga Chlamydomonas reinhardtii.

作者信息

Neupert Juliane, Shao Ning, Lu Yinghong, Bock Ralph

机构信息

Max-Planck-Institut für Molekulare Pflanzenphysiologie, Potsdam-Golm, Germany.

出版信息

Methods Mol Biol. 2012;847:35-47. doi: 10.1007/978-1-61779-558-9_4.

Abstract

Over the past three decades, the single-celled green alga Chlamydomonas reinhardtii has become an invaluable model organism in plant biology and an attractive production host in biotechnology. The genetic transformation of Chlamydomonas is relatively simple and efficient, but achieving high expression levels of foreign genes has remained challenging. Here, we provide working protocols for algal cultivation and transformation as well as for selection and analysis of transgenic algal clones. We focus on two commonly used transformation methods for Chlamydomonas: glass bead-assisted transformation and particle gun-mediated (biolistic) transformation. In addition, we describe available tools for promoting efficient transgene expression and highlight important considerations for designing transformation vectors.

摘要

在过去三十年中,单细胞绿藻莱茵衣藻已成为植物生物学中一种极具价值的模式生物,也是生物技术领域颇具吸引力的生产宿主。莱茵衣藻的遗传转化相对简单高效,但实现外源基因的高表达水平仍具有挑战性。在此,我们提供了藻类培养与转化以及转基因藻类克隆筛选与分析的操作方案。我们重点介绍莱茵衣藻两种常用的转化方法:玻璃珠辅助转化和粒子枪介导(生物弹道)转化。此外,我们还描述了促进转基因高效表达的可用工具,并强调了设计转化载体时的重要注意事项。

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