Abstract：
Developing microorganisms with a high ribonucleic acid (RNA) content is crucial for the RNA industry. Numerous studies have been conducted to enhance RNA production in yeast cells through genetic engineering, yet precise mechanisms remain elusive. Previously, upregulation of TAL1 or PGM2 and deleting PRS5 or DBP8 individually could increase the RNA content in Saccharomyces pastorianus. In this study, within these genetically modified strains, the intracellular nucleotide levels notably increased following cell fragmentation. Deletion of PRS5 and DBP8 within the strain prompted the upregulation of genes sharing similar functions, consequently augmenting the flow of the gene pathway. Furthermore, the upregulation of genes encoding cell-cycle-dependent protein kinases (CDK) was observed in the G03-△PRS5 strain. The influence of TAL1 and PGM2 on RNA content was attributed to the pentose phosphate pathway (PPP). The RNA content of polygenic recombinant strains, G03-△PRS5+△DBP8 and G03-△PRS5+△DBP8+PGM2, displayed the most significant improvement, increasing by 71.8 and 80.1% when compared to the parental strain. Additionally, the maximum specific growth rate of cells increased in these strains. This study contributes valuable insights into the genetic mechanisms underlying high nucleic acid synthesis in S. pastorianus.
摘要：
开发具有高核糖核酸(RNA)含量的微生物对于RNA工业至关重要。已经进行了许多研究以通过基因工程增强酵母细胞中的RNA产生,然而精确的机制仍然难以捉摸。以前,TAL1或PGM2的上调和PRS5或DBP8的单独删除可以增加巴氏酵母中的RNA含量。在这项研究中,在这些转基因菌株中，细胞断裂后，细胞内核苷酸水平显着增加。菌株中PRS5和DBP8的缺失促使具有相似功能的基因上调,从而增加了基因通路的流动。此外，在G03-△PRS5菌株中观察到编码细胞周期依赖性蛋白激酶（CDK）的基因上调。TAL1和PGM2对RNA含量的影响归因于戊糖磷酸途径(PPP)。多基因重组菌株的RNA含量,G03-△PRS5+△DBP8和G03-△PRS5+△DBP8+PGM2，表现出最显著的改善，与亲本菌株相比，分别增加了71.8%和80.1%。此外,在这些菌株中，细胞的最大比生长速率增加。这项研究为巴斯德氏链球菌高核酸合成的遗传机制提供了有价值的见解。
