Selectable Markers and Reporter Genes for Engineering the Chloroplast of Chlamydomonas reinhardtii.
Selectable Markers and Reporter Genes for Engineering the Chloroplast of Chlamydomonas reinhardtii.
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DOI：
10.3390/biology7040046
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发表时间：
2018-10-10
期刊：
影响因子：
4.2
通讯作者：
Purton S
中科院分区：
文献类型：
--
作者：
Esland L;Larrea-Alvarez M;Purton S
莱茵衣藻（Chlamydomonas reinhardtii）是一种模式藻，作为生产有价值化合物（包括治疗性蛋白质和生物活性代谢物）的细胞工厂，越来越受到人们的关注。外源基因在叶绿体中的表达是特别有利的，因为：(i)在这个亚细胞区室中积累的产物将对细胞其余部分的潜在毒性降到最低；（ii）基因可以通过同源重组整合到叶绿体基因组（质体）的特定位点；（iii）利用质体的高倍性和叶绿体基因的高表达，可以获得高达细胞总蛋白5%的重组蛋白水平；（iv）叶绿体中缺乏任何基因沉默机制，确保了转基因的稳定表达。然而，莱茵青霉叶绿体转化体的产生需要有效的选择方法，以及随后标记去除的理想方法。此外，报告基因的使用对于实现对基因表达的全面理解至关重要，从而为重组应用的实验设计提供信息。本文综述了目前可用于莱茵苔叶绿体工程的选择和报告系统，以及用于高等植物和其他微藻叶绿体工程的报告系统，并展望了未来可能出现的新的标记和报告系统，这些标记和报告系统将进一步推动莱茵苔叶绿体作为表达平台的发展。
Chlamydomonas reinhardtii is a model alga of increasing interest as a cell factory for the production of valuable compounds, including therapeutic proteins and bioactive metabolites. Expression of foreign genes in the chloroplast is particularly advantageous as: (i) accumulation of product in this sub-cellular compartment minimises potential toxicity to the rest of the cell; (ii) genes can integrate at specific loci of the chloroplast genome (plastome) by homologous recombination; (iii) the high ploidy of the plastome and the high-level expression of chloroplast genes can be exploited to achieve levels of recombinant protein as high as 5% total cell protein; (iv) the lack of any gene silencing mechanisms in the chloroplast ensures stable expression of transgenes. However, the generation of C. reinhardtii chloroplast transformants requires efficient methods of selection, and ideally methods for subsequent marker removal. Additionally, the use of reporter genes is critical to achieving a comprehensive understanding of gene expression, thereby informing experimental design for recombinant applications. This review discusses currently available selection and reporter systems for chloroplast engineering in C. reinhardtii, as well as those used for chloroplast engineering in higher plants and other microalgae, and looks to the future in terms of possible new markers and reporters that will further advance the C. reinhardtii chloroplast as an expression platform.
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影响因子：
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发表时间：
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期刊：
MOLECULAR AND GENERAL GENETICS
影响因子：
--
作者：
Bateman, JM;Purton, S
通讯作者：
Purton, S