通过诱导功能性ncRNA-SRG1的高水平转录在酿酒酵母中高效产生RNA。

IF 4.1 2区 生物学 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Can Guo , Zhiqiang Bin , Pengjie Zhang , Jing Tang , Lianqing Wang , Yefu Chen , Dongguang Xiao , Xuewu Guo
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引用次数: 0

摘要

RNA(核糖核酸)是生物体的重要组成部分,广泛应用于食品和制药工业。酿酒酵母菌是公认的安全菌株,被广泛用于RNA生产。本研究以酿酒葡萄球菌W303-1a为起始菌株,对功能性ncRNA-SRG1进行分子修饰,评价其对RNA生成的影响。同时,过表达其转录相关辅助基因Spt2、Spt6和Cha4,并在培养基中添加丝氨酸诱导SRG1转录,提高SRG1转录水平,研究其对细胞内RNA水平的影响。结果表明,重组菌株W303-1a- srg1的细胞内RNA含量为10.27%,比起始菌株W303-1a的细胞内RNA含量为9.24%提高了11.15%。在此基础上,构建了基因共过表达菌株w303 -1a- srg1 - spt6。同时,采用添加2%丝氨酸策略提高重组菌株SRG1的转录水平和RNA含量。重组菌株的胞内RNA达到11.41%,较起始菌株(W303-1a,未添加丝氨酸)增加23.38%。此外,通过测定菌株中SRG1的转录水平并绘制生长曲线来评估菌株的生长性能。因此,我们发现提高ncRNA的转录水平可以作为构建高RNA含量酿酒葡萄球菌的新思路,为后续相关领域的研究提供有力帮助。本研究为提高酿酒酵母核酸含量提供了一种新的策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Efficient production of RNA in Saccharomyces cerevisiae through inducing high level transcription of functional ncRNA-SRG1
RNA (Ribonucleic Acid) is an essential component of organisms and is widely used in the food and pharmaceutical industries. Saccharomyces cerevisiae, recognized as a safe strain, is widely used for RNA production. In this study, the S. cerevisiae W303–1a was used as a starting strain and molecular modifications were made to the functional ncRNA-SRG1 to evaluate the effect on RNA production. At the same time, its transcriptionally associated helper genes (Spt2, Spt6 and Cha4) were overexpressed and the culture medium was supplemented with serine to induce SRG1 transcription, to increase SRG1 transcription levels and investigate its effect on intracellular RNA levels. The results showed that the intracellular RNA content of the recombinant strain W303–1a-SRG1 was 10.27 %, an increase of 11.15 % compared to the starting strain (W303–1a, with an intracellular RNA content of 9.24 %). On this basis, a gene co-overexpression strain-W303–1a-SRG1-Spt6 was constructed. Simultaneously, the addition of 2 % serine strategy was used to increase the transcription level of SRG1 and RNA content of the recombinant strain. The intracellular RNA of the recombinant strain reached 11.41 %, an increase of 23.38 % compared to the starting strain (W303–1a, without serine supplementation). In addition, the growth performance of the strain was assessed by measuring the SRG1 transcription level in the strain and plotting the growth curve. Therefore, we found that improving the transcription level of ncRNA can be used as a new idea to construct S. cerevisiae with high RNA content, which provides a strong help for subsequent research in related fields. This work provides a new strategy for increasing the nucleic acid content of S. cerevisiae.
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来源期刊
Journal of biotechnology
Journal of biotechnology 工程技术-生物工程与应用微生物
CiteScore
8.90
自引率
2.40%
发文量
190
审稿时长
45 days
期刊介绍: The Journal of Biotechnology has an open access mirror journal, the Journal of Biotechnology: X, sharing the same aims and scope, editorial team, submission system and rigorous peer review. The Journal provides a medium for the rapid publication of both full-length articles and short communications on novel and innovative aspects of biotechnology. The Journal will accept papers ranging from genetic or molecular biological positions to those covering biochemical, chemical or bioprocess engineering aspects as well as computer application of new software concepts, provided that in each case the material is directly relevant to biotechnological systems. Papers presenting information of a multidisciplinary nature that would not be suitable for publication in a journal devoted to a single discipline, are particularly welcome.
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