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the plant journal:最优密码子设计增强绿藻中的外源基因表达

已有 144 次阅读 2018-3-13 08:59 |个人分类:每日摘要|系统分类:论文交流


Enhancing heterologous expression in Chlamydomonas reinhardtii by transcript sequence optimization

First author: Iddo Weiner; Affiliations: Tel Aviv University (特拉维夫大学): Tel Aviv, Israel

Corresponding author: Iftach Yacoby

Various species of microalgae (微藻) have recently emerged as promising host-organisms (宿主生物) for use in biotechnology industries due to their unique properties. These include efficient conversion of sunlight into organic compounds (有机化合物), the ability to grow in extreme conditions and the occurrence of numerous post-translational (翻译后) modification pathways. However, the inability to obtain high levels of nuclear heterologous (异源的) gene expression in microalgae hinders (阻碍) the development of the entire field. To overcome this limitation, we analyzed different sequence optimization algorithms while studying the effect of transcript sequence features on heterologous expression in the model microalga Chlamydomonas reinhardtii (莱茵藻), whose genome consists of rare features such as a high GC content. Based on the analysis of genomic data, we created eight unique sequences coding for a synthetic ferredoxin–hydrogenase enzyme (铁氧还蛋白氢化酶), used here as a reporter gene. Following in silico (生物信息学) design, these synthetic genes were transformed into the C. reinhardtii nucleus, after which gene expression levels were measured. The empirical data (实验数据), measured in vivo show a discrepancy (差异) of up to 65-fold between the different constructs. In this work we demonstrate how the combination of computational methods and our empirical results enable us to learn about the way gene expression is encoded in the C. reinhardtii transcripts. We describe the deleterious effect on overall expression of codons encoding for splicing signals. Subsequently, our analysis shows that utilization of a frequent subset of preferred codons results in elevated transcript levels, and that mRNA folding energy in the vicinity (附近) of translation initiation (翻译起始) significantly affects gene expression.


通讯Iftach Yacoby (https://en-lifesci.tau.ac.il/profile/iftachy)



doi: 10.1111/tpj.13836

Journal: the plant journal

Published date: 12 March, 2018

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