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Mutagenic effect of ultraviolet radiation and trimethyl psoralen in Mycoplasma towards minimal genome. 紫外线辐射和三甲基补骨脂素对支原体最小基因组的诱变作用
IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-08-10 DOI: 10.1266/ggs.24-00061
Kaito Seo, Kensei Okada, Norikazu Ichihashi

Mycoplasma, solely culturable bacteria with the smallest genome, is an important organism to understand the minimal form of life. Mutagenesis using mutagens is a useful methodology for understanding the essential regions of genomic information. Ultraviolet light and trimethyl psoralen are mutagens known to induce various mutations; the latter is reported to specifically induce deletions in nematodes. However, their mutagenic effects on mycoplasma are not known. Here, we exposed Metamycoplasma salivarium to ultraviolet (UV) light or trimethyl psoralen and UV as mutagens, and analyzed the mutational pattern after several rounds of serial cultivation ranging from 34 to 56 for different lineages. Our results showed that more deletions, but fewer point mutations, were induced with TMP and UV-A than with UV alone, indicating the usefulness of TMP in inducing deletions. In addition, we compared our results with mutational data from other studies, which suggested that both TMP-UVA and UV exposure induced point mutations that were highly biased toward C to T and G to A transitions. These data provide useful basic knowledge for mutational studies on M. salivarium.

支原体是基因组最小的唯一可培养细菌,是了解生命最小形式的重要生物。使用诱变剂进行诱变是了解基因组信息重要区域的有效方法。紫外线和三甲基补骨脂素是已知可诱导各种突变的诱变剂;据报道,后者可特异性地诱导线虫体内的缺失。然而,它们对支原体的诱变作用尚不清楚。在此,我们将唾液支原体暴露于紫外线(UV)或三甲基补骨脂素和紫外线作为诱变剂,并分析了不同品系经过 34 至 56 轮连续培养后的突变模式。我们的结果表明,与单独使用紫外线相比,使用 TMP 和 UV-A 诱导的缺失更多,但点突变更少,这表明 TMP 有助于诱导缺失。此外,我们还将我们的结果与其他研究的突变数据进行了比较,结果表明 TMP-UVA 和紫外线照射诱导的点突变高度偏向于 C 到 T 和 G 到 A 的转变。这些数据为唾液腺霉菌的突变研究提供了有用的基础知识。
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引用次数: 0
Latitudinal adaptation and dispersal pathway of foxtail millet suggested by geographical distribution of transposable elements inserted in the SiPRR37 gene. 插入狐尾粟 SiPRR37 基因的转座元件的地理分布所提示的纬度适应性和扩散途径。
IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-25 Epub Date: 2024-05-24 DOI: 10.1266/ggs.24-00023
Kenji Fukunaga, Akira Abe, Kazue Ito, Kaori Oikawa, Masaya Tsuji, Makoto Kawase

We investigated the variation and geographical distribution of the Pseudo-regulator response 37 (Setaria italica PRR37; SiPRR37) gene, which is involved in heading time (photoperiodism) in foxtail millet. An allele of the SiPRR37 gene, in which an approximately 4.9-kb transposable element (designated TE1) is inserted (a loss-of-function or reduction-of-function type), is distributed sporadically in East Asia and broadly in Southeast Asia and South Asia, implying that this gene is important in latitudinal adaptation. In addition, we found a new allele of SiPRR37 with an insertion of a 360-bp TE (TE2) at this locus and investigated the geographical distribution of this new type. This SiPRR37 allele with TE2 is distributed in Japan, Korea, Nepal, Iran and Turkey. Both TE1 and TE2 are useful markers for tracing foxtail millet dispersal pathways in Asia.

我们研究了狐尾黍中参与顶端时间(光周期)的伪调节器反应 37(Setaria italica PRR37=SiPRR37)基因的变异和地理分布。SiPRR37基因的一个等位基因插入了一个约4.9kb的转座元件(本文中称为TE1)(功能缺失型或功能降低型),该等位基因零星分布于东亚,广泛分布于东南亚和南亚,这意味着该基因在纬度适应方面具有重要作用。此外,我们还在该基因位点上发现了一个插入了 360-bp TE(被命名为 TE2)的 SiPRR37 新等位基因,并对这一新类型的地理分布进行了研究。这种带有 TE2 的 SiPRR37 等位基因分布在日本、韩国、尼泊尔、伊朗和土耳其。TE1 和 TE2 都是追踪狐尾粟在亚洲扩散路径的有用标记。
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引用次数: 0
β-Tyrosine and its biosynthetic enzyme TAM1 are predominantly distributed in the ancestral subpopulation of japonica rice in Oryza rufipogon. β-酪氨酸及其生物合成酶 TAM1 主要分布在粳稻的祖先亚群 Oryza rufipogon 中。
IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-19 DOI: 10.1266/ggs.24-00017
Shunta Sakamoto, Takanori Yoshikawa, Yutaka Sato, Naoki Mori

Intraspecific variations in specialized metabolites play a crucial role in the adaptive response to diverse environments. Two major subspecies, japonica and indica, are observed in Asian cultivated rice (Oryza sativa L.). Previously, we identified (3R)-β-tyrosine, a novel nonproteinogenic β-amino acid in plants, along with the enzyme tyrosine aminomutase (TAM1), required for β-tyrosine biosynthesis, in the japonica cultivar Nipponbare. Notably, TAM1 and β-tyrosine preferentially distributed in japonica cultivars compared with indica cultivars. Considering its phytotoxicity and antimicrobial activity, intraspecific variations in β-tyrosine may contribute to defensive potentials of japonica rice. Investigation of the evolutionary trajectory of TAM1 and β-tyrosine should enhance our understanding of evolution of rice defense. However, their distribution patterns in Oryza rufipogon, the direct ancestor of O. sativa, remain unclear. Therefore, in this study, we extensively examined TAM1 presence/absence and β-tyrosine contents involving 110 genetically and geographically diverse O. rufipogon and revealed that they are characteristically observed in the ancestral subpopulation of japonica rice, while being absent or slightly accumulated in other subpopulations. Thus, we conclude that TAM1 and β-tyrosine in japonica rice are likely derived from its ancestral subpopulation. Furthermore, the high and low TAM1 possession rates and β-tyrosine contents in japonica and indica rice, respectively, could be attributed to distribution patterns of TAM1 and β-tyrosine in their ancestral subpopulations. This study provides fundamental insights into evolution of rice defense.

特化代谢物的种内差异在对不同环境的适应性反应中起着至关重要的作用。亚洲栽培稻(Oryza sativa L.)有两个主要亚种,即粳稻和籼稻。此前,我们在粳稻栽培品种 Nipponbare 中发现了植物中一种新的非蛋白原性 β-氨基酸 (3R)-β-酪氨酸,以及 β-酪氨酸生物合成所需的酪氨酸氨化酶 (TAM1)。值得注意的是,与籼稻栽培品种相比,TAM1 和 β-酪氨酸在粳稻栽培品种中分布更多。考虑到其植物毒性和抗菌活性,β-酪氨酸的种内差异可能有助于提高粳稻的防御潜力。对 TAM1 和 β-酪氨酸进化轨迹的研究应能加深我们对水稻防御进化的理解。然而,它们在 Oryza rufipogon(O. sativa 的直系祖先)中的分布模式仍不清楚。因此,在本研究中,我们广泛研究了 TAM1 的存在/缺失和 β-酪氨酸的含量,涉及 110 个具有遗传和地理多样性的 Oryza rufipogon,结果发现它们在粳稻的祖先亚群中具有典型的观察特征,而在其他亚群中缺失或轻微积累。因此,我们得出结论,粳稻中的 TAM1 和 β-酪氨酸很可能来自其祖先亚群。此外,粳稻和籼稻中 TAM1 的拥有率和 β-酪氨酸含量的高低可能与 TAM1 和 β-酪氨酸在其祖先亚群中的分布模式有关。这项研究为水稻的防御进化提供了基本见解。
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引用次数: 0
Identification of abiotic stress responsive genes: A genome wide analysis of the cytokinin response regulator gene family in rice. 鉴定非生物胁迫响应基因:水稻细胞分裂素响应调节基因家族的全基因组分析
IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-29 DOI: 10.1266/ggs.24-00068
Setu Rani Saha, S M Shahinul Islam, Kimiko Itoh

Response regulators (RRs), which are implicated in various developmental processes as well as environmental responses by acting as either positive or negative regulators, are crucial components of cytokinin signaling in plants. We characterized 36 RRs using in silico and computational analyses of publicly available data. A comprehensive analysis of OsRR family members was performed covering their physicochemical properties, chromosomal distribution, subcellular localization, phylogeny, gene structure, distribution of conserved motifs and domains, and gene duplication events. Gene Ontology analysis results indicate that 22 OsRR genes contribute mainly to the cytokinin-response and signal transduction. Predicted cis-elements in RRs promoter sequences related to phytohormones and abiotic stresses indicate that RRs are involved in hormonal and environmental responses as described in previous studies. MicroRNA (miRNA) target analysis showed that 148 miRNAs target 29 OsRR genes. In some cases, those RRs are targets of the same miRNA group, and may be controlled by common stimulus responses. Based on the analysis of publicly available gene expression data, OsRR4, OsRR6, OsRR9, OsRR10, OsRR22, OsPRR73, and OsPRR95 were found to be involved in response to abiotic stresses. Using quantitative reverse transcription polymerase chain reaction (qPCR) we confirmed that those RRs, namely OsRR4, OsRR6, OsRR9, OsRR10, OsRR22, and OsPRR73, are involved in the response to salinity, osmotic, alkaline and wounding stresses, and can potentially be used as models to understand molecular mechanisms underlying stress responsiveness.

反应调节因子(RRs)通过作为正性或负性调节因子参与各种发育过程和环境反应,是植物细胞分裂素信号转导的关键组成部分。我们通过对公开数据进行硅学和计算分析,确定了 36 个 RRs 的特征。我们对 OsRR 家族成员进行了全面的分析,包括其理化性质、染色体分布、亚细胞定位、系统发育、基因结构、保守基序和结构域的分布以及基因复制事件。基因本体分析结果表明,22 个 OsRR 基因主要参与细胞分裂素反应和信号转导。RRs启动子序列中与植物激素和非生物胁迫相关的顺式元件预测表明,RRs参与了激素和环境响应,这在之前的研究中已有描述。微RNA(miRNA)靶标分析表明,148个miRNA以29个OsRR基因为靶标。在某些情况下,这些 RR 是同一 miRNA 组的靶标,可能受共同刺激反应的控制。根据对公开基因表达数据的分析,发现 OsRR4、OsRR6、OsRR9、OsRR10、OsRR22、OsPRR73 和 OsPRR95 参与了对非生物胁迫的响应。利用定量反转录聚合酶链式反应(qPCR),我们证实了这些 RRs,即 OsRR4、OsRR6、OsRR9、OsRR10、OsRR22 和 OsPRR73,参与了对盐度、渗透压、碱性和伤口胁迫的响应,并有可能被用作了解胁迫响应分子机制的模型。
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引用次数: 0
Development of polymorphic microsatellite markers for distylous-homostylous Primula secundiflora (Primulaceae) using HiSeq sequencing. 利用 HiSeq 测序技术开发多态性微卫星标记,用于二歧同株报春花(报春花科)。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2024-05-24 Epub Date: 2024-04-13 DOI: 10.1266/ggs.23-00340
Hua-Ying Sun, Wen-Ping Zhang, Wei Zhou, Zhi-Kun Wu, Lan-Ping Zheng

Primula secundiflora is an insect-pollinated, perennial herb belonging to the section Proliferae (Primulaceae) that exhibits considerable variation in its mating system, with predominantly outcrossing populations comprising long-styled and short-styled floral morphs and selfing populations comprising only homostyles. To facilitate future investigations of the population genetics and mating patterns of this species, we developed 25 microsatellite markers from P. secundiflora using next-generation sequencing and measured polymorphism and genetic diversity in a sample of 30 individuals from three natural populations. The markers displayed high polymorphism, with the number of observed alleles per locus ranging from three to 16 (mean = 8.36). The observed and expected heterozygosities ranged from 0.100 to 1.000 and 0.145 to 0.843, respectively. Twenty-one of the loci were also successfully amplified in P. denticulata. These microsatellite markers should provide powerful tools for investigating patterns of population genetic diversity and the evolutionary relationships between distyly and homostyly in P. secundiflora.

报春花(Primula secundiflora)是一种昆虫授粉的多年生草本植物,属于报春花科(Proliferae),在交配系统方面表现出相当大的差异,主要是由长型和短型花形态组成的外交种群和仅由同型花组成的自交种群。为便于今后对该物种的种群遗传学和交配模式进行研究,我们利用新一代测序技术开发了 25 个 P. secundiflora 的微卫星标记,并测量了来自三个自然种群的 30 个个体的多态性和遗传多样性。标记显示出相对较高的多态性,每个位点观察到的等位基因数量从 3 个到 16 个不等(平均值 = 8.36)。观察到的杂合性和预期杂合性分别为 0.100 至 1.000 和 0.145 至 0.843。其中 21 个位点也在 P. denticulata 中成功扩增。这些微卫星标记可为研究该物种的种群遗传多样性模式以及畸形和同形之间的进化关系提供有力的工具。
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引用次数: 0
METTL21C mediates autophagy and formation of slow-twitch muscle fibers in mice after exercise. METTL21C 在运动后介导自噬的发生和慢肌纤维的形成。
IF 1 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-05-10 Epub Date: 2024-04-18 DOI: 10.1266/ggs.23-00320
Jing Qu, Shuai Dang, Yuan-Yuan Sun, Tao Zhang, Hai Jiang, Hong-Zhao Lu

Homeostasis is essential for muscle repair and regeneration after skeletal muscle exercise. This study investigated the role of methyltransferase-like 21C (METTL21C) in skeletal muscle of mice after exercise and the potential mechanism. First, muscle samples were collected at 2, 4 and 6 weeks after exercise, and liver glycogen, muscle glycogen, blood lactic acid and triglyceride were assessed. Moreover, the expression levels of autophagy markers and METTL21C in skeletal muscle were analyzed. The results showed that the expression levels of METTL21C and MYH7 in the gastrocnemius muscle of mice in the exercise group were significantly higher after exercise than those in the control group, which suggested that long-term exercise promoted the formation of slow-twitch muscle fibers in mouse skeletal muscle. Likewise, the autophagy capacity was enhanced with the prolongation of exercise in muscles. The findings were confirmed in mouse C2C12 cells. We discovered that knockdown of Mettl21c reduced the expression of MYH7 and the autophagy level in mouse myoblasts. These findings indicate that METTL21C promotes skeletal muscle homeostasis after exercise by enhancing autophagy, and also contributes to myogenic differentiation and the formation of slow muscle fibers.

骨骼肌运动后的肌肉修复和再生离不开平衡。本研究探讨了甲基转移酶样 21C(METTL21C)在小鼠运动后骨骼肌中的作用及其潜在机制。首先,在运动后2周、4周和6周采集肌肉样本,评估肝糖原、肌糖原、血乳酸(BLA)和甘油三酯(TG)。此外,还分析了骨骼肌中自噬标记物和 METTL21C 的表达水平。结果显示,运动组小鼠运动后腓肠肌中METTL21C和MYH7的表达量明显高于对照组,这表明长期运动促进了小鼠骨骼肌中慢肌纤维的形成。同样,肌肉的自噬能力也会随着运动时间的延长而增强。这些发现在小鼠 C2C12 细胞中得到了进一步验证。我们发现,敲除 Mettl21c 会降低小鼠肌母细胞中 MYH7 的表达和自噬水平。这些发现表明,METTL21C通过增强自噬促进了运动后骨骼肌的稳态,同时METTL21C还有助于肌原纤维的分化和慢肌纤维的形成。
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引用次数: 0
A budding yeast CAGE dataset comprising two cell types. 芽殖酵母帽分析两种细胞类型的基因表达数据集。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2024-04-18 Epub Date: 2024-04-13 DOI: 10.1266/ggs.24-00020
Kei Kawakami, Shin-Ichi Maeda, Yoshiko Tanimoto, Mitsuhiro Shimizu, Hiroaki Kato

The budding yeast Saccharomyces cerevisiae is an excellent model organism for studying chromatin regulation with high-resolution genome-wide analyses. Since newly generated genome-wide data are often compared with publicly available datasets, expanding our dataset repertoire will be beneficial for the field. Information on transcription start sites (TSSs) determined at base pair resolution is essential for elucidating mechanisms of transcription and related chromatin regulation, yet no datasets that cover two different cell types are available. Here, we present a CAGE (cap analysis of gene expression) dataset for a-cells and α-cells grown in defined and rich media. Cell type-specific genes were differentially expressed as expected, ensuring the reliability of the data. Some of the differentially expressed TSSs were medium-specific or detected due to unrecognized chromosome rearrangement. By comparing the CAGE data with a high-resolution nucleosome map, major TSSs were primarily found in +1 nucleosomes, with a peak approximately 30 bp from the promoter-proximal end of the nucleosome. The dataset is available at DDBJ/GEA.

芽殖酵母是利用高分辨率全基因组分析研究染色质调控的极佳模式生物。由于新生成的全基因组数据经常与公开数据集进行比较,因此扩大我们的数据集范围将有利于该领域的研究。以碱基对分辨率测定的转录起始位点(TSS)信息对于阐明转录和相关染色质调控机制至关重要,但目前还没有涵盖两种不同细胞类型的数据集。在这里,我们展示了在限定培养基和富集培养基中生长的 a 细胞和 α 细胞的基因表达上限分析(CAGE)数据集。细胞类型特异基因的差异表达符合预期,确保了数据的可靠性。一些差异表达的 TSS 具有培养基特异性,或由于未识别的染色体重排而被检测到。通过将 CAGE 数据与高分辨率核小体图进行比较,主要的 TSSs 主要存在于 +1 核小体中,其峰值距离核小体的启动子近端约 30 bp。该数据集可在 DDBJ/GEA 网站上查阅。
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引用次数: 0
Current research on mechanisms of limb bud development, and challenges for the next decade. 目前关于肢芽发育机制的研究;未来十年面临的困难。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2024-03-29 Epub Date: 2024-02-21 DOI: 10.1266/ggs.23-00287
Takayuki Suzuki

The developmental mechanisms of limb buds have been studied in developmental biology as an excellent model of pattern formation. Chick embryos have contributed to the discovery of new principles in developmental biology, as it is easy to observe live embryos and manipulate embryonic tissues. Herein, I outline recent findings and future issues over the next decade regarding three themes, based on my research: limb positioning, proximal-distal limb elongation and digit identity determination. First, how hindlimb position is determined at the molecular level is described, with a focus on the transforming growth factor-β signaling molecule GDF11. Second, I explain how the cell population in the limb bud deforms with developmental progress, shaping the limb bud with elongation along the proximal-distal axis. Finally, I describe the developmental mechanisms that determine digit identity through the interdigits.

在发育生物学中,肢芽的发育机制一直是研究模式形成的一个重要模型。雏鸡胚胎易于观察活体胚胎和操作胚胎组织,因此为发现发育生物学的新原理做出了贡献。在此,我们根据自己的研究,概述了目前的发现和未来 10 年的三个主题:1)肢体定位;2)肢体近端-远端伸长;3)数字身份确定。首先,我们以转化生长因子-β(TGF-β)信号分子 GDF11 为中心,阐述了后肢位置是如何在分子水平上决定的。其次,我们解释了肢芽中的细胞群如何随着发育进程而变形,从而形成沿近端-远端轴伸长的肢芽。最后,我们描述了通过指间体决定指位特征的发育机制。
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引用次数: 0
Pigmentation of soybean seed coats via a mutation that abolishes production of multiple-phased siRNAs of chalcone synthase genes. 大豆种皮的色素沉着是通过抑制查尔酮合成酶基因多相 siRNA 的突变产生的。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2024-03-29 Epub Date: 2024-02-21 DOI: 10.1266/ggs.23-00260
Mashiro Yuhazu, Shun Mikuriya, Ayumi Mori, Maria Stefanie Dwiyanti, Mineo Senda, Akira Kanazawa

Lack of pigmentation in seed coats of soybean is caused by natural RNA silencing of chalcone synthase (CHS) genes. This phenomenon is an evolutionary consequence of structural changes in DNA that resulted in the production of double-stranded RNAs (dsRNAs) that trigger RNA degradation. Here we determined that a mutant with pigmented seed coats derived from a cultivar that lacked the pigmentation had a deletion between DNA regions ICHS1 and a cytochrome P450 gene; the deletion included GmIRCHS, a candidate gene that triggers CHS RNA silencing via production of CHS dsRNAs. We also characterized CHS short interfering RNAs (siRNAs) produced in the wild-type seed coats that had CHS RNA silencing. Phased 21-nt CHS siRNAs were detected in all 21 phases and were widely distributed in exon 2 of CHS7, which indicates commonality in the pattern of RNA degradation in natural CHS RNA silencing between distantly related species. These results with the similarities in the rearrangements found in spontaneous mutants suggest that the structural organization that generates dsRNAs that trigger phased siRNA production is vulnerable to further structural changes, which eventually abolish the induction of RNA silencing.

大豆种皮缺乏色素是由查尔酮合成酶(CHS)基因的天然 RNA 沉默引起的。这种现象是 DNA 结构变化导致双链 RNA(dsRNA)产生并引发 RNA 降解的进化结果。在这里,我们确定了一个从缺乏色素的栽培品种中获得的具有色素种皮的突变体,其 DNA 区域 ICHS1 和细胞色素 P450 基因之间存在缺失;该缺失包括 GmIRCHS,这是一个通过产生 CHS dsRNAs 触发 CHS RNA 沉默的候选基因。我们还对野生型种皮中产生的 CHS 短干扰 RNA(siRNA)进行了鉴定,这些短干扰 RNA 具有 CHS RNA 沉默功能。在所有 21 个阶段中都检测到了分阶段的 21-nt CHS siRNA,它们广泛分布在 CHS7 的第 2 外显子中,这表明在远缘物种之间的天然 CHS RNA 沉默中,RNA 降解模式具有共性。这些结果与在自发突变体中发现的重排相似,表明产生 dsRNAs 触发阶段性 siRNA 产生的结构组织很容易发生进一步的结构变化,最终导致 RNA 沉默的诱导失效。
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引用次数: 0
RNA synthesis in liposomes with negatively charged lipids after fusion via freezing-thawing. 带负电荷的脂质体通过冷冻-解冻融合后的 RNA 合成。
IF 1.1 4区 生物学 Q2 Medicine Pub Date : 2024-03-26 Epub Date: 2024-02-21 DOI: 10.1266/ggs.23-00297
Gakushi Tsuji, Ayu Shimomura, Shota Fukuoka, Masaya Oki

The freezing-thawing (F/T) method for fusing giant unilamellar vesicles (GUVs) can provide substrates, enzymes and membrane material simultaneously and repetitively, and is useful for constructing a recursive model of an artificial cell. However, enzymatic efficiency after F/T is reduced due to rupture of the GUVs and leakage of the inner solution during F/T. Previously, liposomes composed of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and a negatively charged lipid, such as 1-palmitoyl-2-oleoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (POPG), showed lower rupture and leakage rates during F/T. In this study, we investigated the effect of POPG on the supply of components required for T7 RNA polymerase reactions via F/T by flow cytometry analysis. We found that the addition of POPG to liposome preparations reduced the efficiency of RNA synthesis. In addition, DNA was concentrated during F/T and RNA synthesis occurred after F/T in liposomes composed of POPC and POPG. Our results provide new insights for more efficient supply of substrates and enzymes by the F/T method, thereby increasing the utility of the F/T method for the construction of recursive bioreactors.

用冷冻-解冻(F/T)方法融合巨型单拉美米尔囊泡(GUVs)可同时重复提供底物、酶和膜材料,有助于构建人工细胞递归模型。然而,在 F/T 过程中,由于 GUV 破裂和内部溶液泄漏,F/T 后的酶效率会降低。以前,由 1-棕榈酰-2-油酰-sn-甘油-3-磷酸胆碱(POPC)和带负电荷的脂质(如 1-棕榈酰-2-油酰-sn-甘油-3-磷酸-(1'-rac-甘油)(POPG)组成的脂质体在 F/T 期间显示出较低的破裂率和泄漏率。在本研究中,我们通过流式细胞仪分析,研究了 POPG 对 T7 RNA 聚合酶反应所需成分供应的影响。我们发现,在脂质体制备过程中加入 POPG 会降低 RNA 合成的效率。此外,在由 POPC 和 POPG 组成的脂质体中,DNA 在 F/T 期间被浓缩,而 RNA 的合成则在 F/T 之后进行。我们的研究结果为利用 F/T 法更有效地提供底物和酶提供了新的启示,从而提高了 F/T 法在构建递归生物反应器方面的实用性。
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引用次数: 0
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