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裂殖酵母中转录和翻译效率之间的相互关系。

Reciprocal relation between reporter gene transcription and translation efficiency in fission yeast.

机构信息

Central Research Institute, Kasauli, Distt, Solan, Himachal Pradesh 173204, India; Department of Molecular Biology, Institute of Microbial Technology, Sector 39A, Chandigarh-160036. India.

Central Research Institute, Kasauli, Distt, Solan, Himachal Pradesh 173204, India; Pharmacology Department, School of Science and Technology, Nottingham Trent University, Nottingha, NG11 8NS, UK.

出版信息

Plasmid. 2021 May;115:102557. doi: 10.1016/j.plasmid.2021.102557. Epub 2021 Feb 2.

Abstract

The fission yeast, Schizosaccharomyces pombe, is an excellent model for basic research but is not useful for commercial scale protein expression due to lack of strong expression vectors. Earlier, we showed that the lsd90 promoter elicited significantly greater GFP expression level than the adh1 and nmt1 promoters, albeit in different vector backbones. Here, we have systematically investigated the contribution of selectable markers, LEU2 and URA3m to GFP expression: while LEU2 elicited very low expression, the URA3m gene, with truncated promoter, elicited much greater GFP expression level with all promoters. Paradoxically, an inverse correlation was observed between the GFP transcription and translation efficiency. This system can be useful for understanding the factors governing recombinant gene expression and optimization of protein production.

摘要

裂殖酵母(Schizosaccharomyces pombe)是基础研究的优秀模型,但由于缺乏强表达载体,因此不适用于商业规模的蛋白质表达。早期,我们发现 lsd90 启动子比 adh1 和 nmt1 启动子引发更高水平的 GFP 表达,尽管在不同的载体骨架中。在这里,我们系统地研究了选择标记物 LEU2 和 URA3m 对 GFP 表达的贡献:虽然 LEU2 引发的表达水平非常低,但具有截短启动子的 URA3m 基因在所有启动子下引发了更高水平的 GFP 表达。矛盾的是,GFP 转录和翻译效率之间观察到负相关。该系统可用于了解调控重组基因表达的因素和优化蛋白质生产。

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