Suppr超能文献

来自[具体来源未给出]的转录激活因子AtxA被用于在[具体对象未给出]中开发一种严格控制、高水平、可调节且特定于稳定期的转录活性。

The transcription activator AtxA from was employed for developing a tight-control, high-level, modulable and stationary-phase-specific transcription activity in .

作者信息

Liu Ying-Xing, Zhuo Xiao-Zhen, Li Si-Yu

机构信息

Department of Chemical Engineering, National Chung Hsing University, Taichung, Taiwan.

Innovation and Development Center of Sustainable Agriculture, National Chung Hsing University, Taichung, Taiwan.

出版信息

Synth Biol (Oxf). 2022 Aug 17;7(1):ysac014. doi: 10.1093/synbio/ysac014. eCollection 2022.

Abstract

The strong transcriptional activity of the virulent gene in has been proven to be anthrax toxin activator (AtxA)-regulated. However, the obscure transcription mechanism hinders practical applications of this strong promoter. In this study, a 509-bp DNA fragment [termed 509sequence, (-508)-(+1) relative to the P2 transcription start site] was cloned upstream of rbs-GFPuv as pTOL02B to elucidate the AtxA-regulated transcription. The 509sequence was dissected into the -10 sequence, -35 sequence, AT tract, SLI/SLII and upstream site. In conjunction with the heterologous co-expression of AtxA (under the control of the T7 promoter), the -10 sequence (TATACT) was sufficient for the AtxA-regulated transcription. Integration of pTOL02F + pTOLAtxA as pTOL03F showed that the AtxA-regulated transcription exhibited a strong specific fluorescence intensity/common analytical chemistry term (OD) of 40 597 ± 446 and an induction/repression ratio of 122. An improved induction/repression ratio of 276 was achieved by cultivating /pTOL03F in M9 minimal medium. The newly developed promoter system termed P consists of AtxA, the -10 sequence and RNA polymerase. These three elements synergistically and cooperatively formed a previously undiscovered transcription system, which exhibited a tight-control, high-level, modulable and stationary-phase-specific transcription. The P was used for expression for the stationary-phase polyhydroxybutyrate production, and the results showed that a PHB yield, content and titer of 0.20 ± 0.27 g/g-glucose, 68 ± 11% and 1.5 ± 0.4 g/l can be obtained. The positive inducible P, in contrast to negative inducible, should be a useful tool to diversify the gene information flow in synthetic biology. Graphical Abstract.

摘要

已证明炭疽杆菌中毒力基因的强转录活性受炭疽毒素激活剂(AtxA)调控。然而,其模糊的转录机制阻碍了这种强启动子的实际应用。在本研究中,将一个509 bp的DNA片段[称为509序列,相对于P2转录起始位点为(-508)-(+1)]克隆到rbs-GFPuv上游,构建成pTOL02B,以阐明AtxA调控的转录。509序列被分解为-10序列、-35序列、AT序列、SLI/SLII和上游位点。与AtxA的异源共表达(在T7启动子控制下)相结合,-10序列(TATACT)足以实现AtxA调控的转录。将pTOL02F + pTOLAtxA整合为pTOL03F表明,AtxA调控的转录表现出40597±446的强特异性荧光强度/共同分析化学术语(OD)和122的诱导/抑制比。通过在M9基本培养基中培养/pTOL03F,诱导/抑制比提高到了276。新开发的称为P的启动子系统由AtxA、-10序列和RNA聚合酶组成。这三个元件协同形成了一个以前未发现的转录系统,该系统表现出严格控制、高水平、可调节和稳定期特异性转录。P被用于稳定期聚羟基丁酸酯生产的表达,结果表明,聚羟基丁酸酯的产量、含量和滴度分别为0.20±0.27 g/g-葡萄糖、68±11%和1.5±0.4 g/l。与负诱导型相比,正诱导型P应该是在合成生物学中使基因信息流多样化的有用工具。图形摘要。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/be0f/9424709/870c70cf767f/ysac014f2.jpg

文献AI研究员

20分钟写一篇综述,助力文献阅读效率提升50倍。

立即体验

用中文搜PubMed

大模型驱动的PubMed中文搜索引擎

马上搜索

文档翻译

学术文献翻译模型,支持多种主流文档格式。

立即体验