{"title":"Reciprocal relation between reporter gene transcription and translation efficiency in fission yeast","authors":"Suchita Srivastava , Satinderdeep Kaur , Hemant K. Verma , Suman Rani , Manisha Thakur , Swati Haldar , Jagmohan Singh","doi":"10.1016/j.plasmid.2021.102557","DOIUrl":null,"url":null,"abstract":"<div><p>The fission yeast, <em>Schizosaccharomyces pombe</em>, 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 <em>lsd90</em> promoter elicited significantly greater GFP expression level than the <em>adh1</em> and <em>nmt1</em> promoters, albeit in different vector backbones. Here, we have systematically investigated the contribution of selectable markers, <em>LEU2</em> and <em>URA3m</em> to GFP expression: while <em>LEU2</em> elicited very low expression, the <em>URA3m</em> 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.</p></div>","PeriodicalId":49689,"journal":{"name":"Plasmid","volume":"115 ","pages":"Article 102557"},"PeriodicalIF":1.8000,"publicationDate":"2021-05-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.plasmid.2021.102557","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Plasmid","FirstCategoryId":"99","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0147619X21000044","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q3","JCRName":"GENETICS & HEREDITY","Score":null,"Total":0}
引用次数: 0
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.
期刊介绍:
Plasmid publishes original research on genetic elements in all kingdoms of life with emphasis on maintenance, transmission and evolution of extrachromosomal elements. Objects of interest include plasmids, bacteriophages, mobile genetic elements, organelle DNA, and genomic and pathogenicity islands.