BMC Biotechnology最新文献

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Correction: Metabolic engineering for microbial production of sugar acids. 更正:微生物生产糖酸的代谢工程。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-06-20 DOI: 10.1186/s12896-025-00996-0
Fatma Gizem Avci, Tim Prasun, Volker F Wendisch
{"title":"Correction: Metabolic engineering for microbial production of sugar acids.","authors":"Fatma Gizem Avci, Tim Prasun, Volker F Wendisch","doi":"10.1186/s12896-025-00996-0","DOIUrl":"10.1186/s12896-025-00996-0","url":null,"abstract":"","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"50"},"PeriodicalIF":3.5,"publicationDate":"2025-06-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12181850/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144336300","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Exploring the mechanisms and mode of action of bioactive compounds from marine Streptomyces albidoflavus against breast carcinoma cells. 探讨海洋白多黄链霉菌生物活性化合物抗乳腺癌的作用机制和方式。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-06-19 DOI: 10.1186/s12896-025-00991-5
Abir M Shata, Mokhtar Saeed Rejili, Manal M El-Naggar, Samy A El-Asser, Ahmed A Saleh, Elsayed E Hafez, Faouzi Haouala, Amany S Youssef
{"title":"Exploring the mechanisms and mode of action of bioactive compounds from marine Streptomyces albidoflavus against breast carcinoma cells.","authors":"Abir M Shata, Mokhtar Saeed Rejili, Manal M El-Naggar, Samy A El-Asser, Ahmed A Saleh, Elsayed E Hafez, Faouzi Haouala, Amany S Youssef","doi":"10.1186/s12896-025-00991-5","DOIUrl":"10.1186/s12896-025-00991-5","url":null,"abstract":"<p><p>Breast cancer remains one of the most lethal diseases for women worldwide. Marine populations are considered a vast reservoir for novel bioactive metabolites, particularly marine Actinomycetes, which are known to produce various bioactive compounds with antitumour, antibacterial, and antifungal properties. A promising new marine strain was isolated and identified as Streptomyces albidoflavus strain EgyAB2 (16 S rRNA gene sequence accession number ON680945.1). The anticancer activity of the extracted compounds was tested in the MCF7 cell line using a sulforhodamine B (SRB) bioassay, which revealed an IC<sub>50</sub> of 0.36 µg/ml compared to the chemotherapeutic drug 5-fluorouracil (0.35 µg/ml). Additionally, the anticancer activity was confirmed by a dimethyl-thiazol-diphenyl-tetrazolium bromide (MTT) bioassay, which showed an IC<sub>50</sub> of 17.46 µg/ml. The mode of action of the treated breast carcinoma (apoptotic effect) was studied via qRT-PCR, revealing a significant role in anticancer treatment. Although the extracted compounds exhibited high antioxidant activity in the diphenyl-2-picrylhydrazyl (DPPH) radical-scavenging assay, they presented an IC<sub>50</sub> of 8.92 µg/ml and an inhibition percentage of 56.08%. Chemical characterisation was performed via GC‒MS, 1 H-NMR, and FTIR spectroscopy analyses, revealing the presence of 2-D N-methyl imidazole, 2-nonadecene, 1-D-2-methyl imidazole, and propane dinitrile, all of which exhibit antitumour activity.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"48"},"PeriodicalIF":3.5,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12180265/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144332411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Newly designed curcumin-loaded hybrid nanoparticles: a multifunctional strategy for combating oxidative stress, inflammation, and infections to accelerate wound healing and tissue regeneration. 新设计的姜黄素负载混合纳米颗粒:对抗氧化应激,炎症和感染的多功能策略,以加速伤口愈合和组织再生。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-06-19 DOI: 10.1186/s12896-025-00989-z
Heidi M Abdel-Mageed, Nermeen Z AbuelEzz, Ahmed A Ali, Amira Emad Abdelaziz, Dina Nada, Sahar M Abdelraouf, Shahinaze A Fouad, Abeer Bishr, Rasha A Radwan
{"title":"Newly designed curcumin-loaded hybrid nanoparticles: a multifunctional strategy for combating oxidative stress, inflammation, and infections to accelerate wound healing and tissue regeneration.","authors":"Heidi M Abdel-Mageed, Nermeen Z AbuelEzz, Ahmed A Ali, Amira Emad Abdelaziz, Dina Nada, Sahar M Abdelraouf, Shahinaze A Fouad, Abeer Bishr, Rasha A Radwan","doi":"10.1186/s12896-025-00989-z","DOIUrl":"10.1186/s12896-025-00989-z","url":null,"abstract":"<p><p>Effective treatment of skin wounds remains a clinical challenge owing to factors such as microbial infections, impaired fibroblast activity, disrupted angiogenesis, and collagen remodeling. In this study, we developed and evaluated novel curcumin-cyclodextrin hybrid nanoparticles (Cur/CD-HNPs) as a multifunctional platform for enhanced wound healing. Nanoparticles (NPs) were prepared via nanoprecipitation. Physicochemical and structural properties were systematically characterized by determining the encapsulation efficiency (EE), particle size, zeta potential, X-XRD, FTIR, SEM, in vitro release, and stability studies. The optimized Cur/CD-HNPs demonstrated a uniform particle size of 150.5 ± 2.8 nm, a surface charge of - 18.5 ± 0.59 mV, a PDI of 0.20 ± 0.03, and a high EE (90.2 ± 2.35%). Cur/CD-HNPs exhibited potent anti-inflammatory effects (97.93 ± 1.24% inhibition of protein denaturation), full antioxidant activity (100% ABTS radical scavenging, IC50 = 12.85 µg/mL), and broad-spectrum antibacterial efficacy. Cur/CD-HNPs exhibited a sustained biphasic release profile, with ~ 82% of Cur released over 24 h, supporting sustained delivery for wound healing applications. In vitro scratch assays revealed enhanced fibroblast proliferation and migration. For in vivo evaluation, the nanoparticles were incorporated into a hydrogel base and applied topically in a rat burn wound model, resulting in significantly accelerated wound closure (P < 0.05). Histopathological examination revealed improved epithelialization, collagen deposition, and tissue regeneration compared with the control groups. Our findings presented Cur/CD-HNPs as a promising therapeutic approach, offering Cur enhanced bioactivity, stability, and regenerative potential. This formulation addresses the key limitations of curcumin and presents a multifunctional and strong translational platform for clinical wound care.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"49"},"PeriodicalIF":3.5,"publicationDate":"2025-06-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12180217/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144332412","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Dietary compounds activates sirtuins - a computational modeling approach towards the management of ageing. 膳食化合物激活sirtuins——一种管理衰老的计算建模方法。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-06-13 DOI: 10.1186/s12896-025-00985-3
Nail Besli, Mazhar T Kaya, Nilufer Ercin, Ulkan Celik
{"title":"Dietary compounds activates sirtuins - a computational modeling approach towards the management of ageing.","authors":"Nail Besli, Mazhar T Kaya, Nilufer Ercin, Ulkan Celik","doi":"10.1186/s12896-025-00985-3","DOIUrl":"10.1186/s12896-025-00985-3","url":null,"abstract":"<p><p>In biogerontology, researchers are actively exploring the manipulation of sirtuin proteins, a family of enzymes that play a crucial role in regulating lifespan and age-related diseases through bioactive compounds found in the diet. This report explores potential new insights by leveraging the innovative intersection of natural bioactive compounds and sirtuin modulation through computational drug design. Using state-of-the-art in silico techniques, including high throughput virtual screening (HTVS) and molecular dynamics (MD) simulation, the study identifies natural compounds from various dietary sources for sirtuin proteins (SIRT 1, 3, 6, 7) related to the cellular process in aging. HTVS was automated with the AutoDock-VINA to pinpoint the novel candidate bioactive compounds for sirtuins, while the MD was conducted with GROMACS. The top hits, fagopyrine, emblicanin B, epicatechin-(4beta- > 8)-epicatechin-(2beta- > 7,4beta- > 8)-catechin (epicatechin), withanolide, and dulcisisoflavone, had superior docking scores ranging from - 12.7 to - 7.2 kcal/mol. The MD displayed steady interactions between target proteins and biocompounds throughout 100 ns without significant conformational shifts. This paper contributes significantly to anti-aging research by presenting a novel, computationally efficient approach to discovering and developing sirtuin-modulating compounds. Highlighting the untapped potential of natural bioactive substances in modulating key biological pathways associated with aging opens new avenues for future experimental exploration of diet-based therapeutic strategies that may support healthy aging.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"47"},"PeriodicalIF":3.5,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12168335/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144309504","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Mycosynthesis of silver nanoparticles by Aspergillus templicola OR480102: a multifaceted approach for antibacterial, anticancer, and scratch assay applications. 由圣殿曲霉OR480102真菌合成纳米银:抗菌,抗癌和划痕试验应用的多方面方法。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-06-11 DOI: 10.1186/s12896-025-00982-6
Marwa M Abdel-Kareem, Maysa M A Ali, Abd El-Latif Hesham, Hossam E F Abdel-Raheam, Marwa Obiedallah
{"title":"Mycosynthesis of silver nanoparticles by Aspergillus templicola OR480102: a multifaceted approach for antibacterial, anticancer, and scratch assay applications.","authors":"Marwa M Abdel-Kareem, Maysa M A Ali, Abd El-Latif Hesham, Hossam E F Abdel-Raheam, Marwa Obiedallah","doi":"10.1186/s12896-025-00982-6","DOIUrl":"10.1186/s12896-025-00982-6","url":null,"abstract":"<p><strong>Background: </strong>Regarding their distinct physico-chemical and bioactivity characteristics, silver nanoparticles 'AgNPs' are extensively utilized in numerous scientific purposes.</p><p><strong>Results: </strong>Within this current investigation, for the first time, we evaluated how the extracellular extract of the isolate MAK223 generated exceptionally fixed AgNPs. The isolate was genetically identified as Aspergillus templicola OR480102. The generated AgNPs' physico-chemical characteristics were assessed using ultraviolet-vis spectroscopy, transmission electron microscopy (TEM), and Fourier transform infrared spectrometry (FT-IR). The maximum absorption in the UV-vis spectrum was obtained at 420 nm, matching the silver nanoparticles' surface plasmon absorbance. A. templicola OR480102 produced uniformly dispersed AgNPs between 5 and 25 nm with a mean dimension of 17.78537 ± 1.36 nm using TEM. FT-IR analysis identified functional groups (e.g., -OH, C = O) in the fungal filtrate that mediate AgNP synthesis and capping. To verify AgNPs stability, the dynamic light scattering (DLS) approach is employed. Optimal conditions for AgNPs synthesis were 10 days of incubation, one mM silver nitrate concentration, pH 11, and elevated temperatures. AgNPs demonstrated efficacy against clinically relevant pathogens: S. typhimurium 'ATCC 14028', B. subtilis 'ATCC 6633', S. aureus 'ATCC 25923', and E. coli 'ATCC 29213' were used in the study. Also, using AgNPs derived from the filtrate of A. templicola OR480102 shows significant potential as a novel therapeutic approach against breast cancer cells 'MCF-7'. The scratch assay of 'MCF-7' cells demonstrates the suppressive impact of AgNPs for these cell lines during proliferation by promoting apoptosis and reducing cell migration.</p><p><strong>Conclusion: </strong>Based on physico-chemical characteristics of AgNPs' and their antimicrobial and anticancer activities, it cleared that the selected strain Aspergillus templicola OR480102 is a promising producer of stable AgNPs' with significant bioactivities which could be applicable in different fields.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"46"},"PeriodicalIF":3.5,"publicationDate":"2025-06-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12153196/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144274058","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Establishment of an indirect ELISA antibody detection method based on the stable expression of LSDV P32 protein in CHO-K1 cells. 建立基于LSDV P32蛋白在CHO-K1细胞中稳定表达的间接ELISA抗体检测方法。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-06-04 DOI: 10.1186/s12896-025-00966-6
Ruolan Xin, Jiawen Zhang, Yinghui Zhang, Daoming Su, Menglin Li, Dan Liu, Ruizhi Wu, Junjie Zhao, Yuanyuan Zhu, Yebing Liu, Xiaoyun Chen, Yongjun Wen, Zhen Zhu
{"title":"Establishment of an indirect ELISA antibody detection method based on the stable expression of LSDV P32 protein in CHO-K1 cells.","authors":"Ruolan Xin, Jiawen Zhang, Yinghui Zhang, Daoming Su, Menglin Li, Dan Liu, Ruizhi Wu, Junjie Zhao, Yuanyuan Zhu, Yebing Liu, Xiaoyun Chen, Yongjun Wen, Zhen Zhu","doi":"10.1186/s12896-025-00966-6","DOIUrl":"10.1186/s12896-025-00966-6","url":null,"abstract":"<p><strong>Background: </strong>Lumpy skin disease (LSD), caused by infection with the lumpy skin disease virus (LSDV), is a highly infectious disease that poses a notable challenge to the cattle industry worldwide. To conduct epidemiological monitoring of LSDV infection in cattle and evaluate the immune efficacy of LSDV vaccines, it is essential to develop a rapid, sensitive, and specific ELISA-based antibody detection method.</p><p><strong>Results: </strong>We utilized the LSDV P32 protein, stably expressed in a CHO-K1 suspension cell system, as a coating antigen to develop an indirect ELISA for Capripoxvirus (CaPV) antibody detection. This method specifically recognizes CaPV-positive sera without cross-reactivity with sera positive for bovine viral diarrhea virus, bovine rotavirus, infectious bovine rhinotracheitis virus, and Brucella antibodies. The method demonstrated a maximum serum dilution detection capacity of 1:3200, with intra- and inter-assay variation coefficients below 10%. Comparison with a commercially available kit showed an agreement of 95.7%.</p><p><strong>Conclusion: </strong>The indirect ELISA antibody detection method established exhibited excellent specificity, sensitivity, and reproducibility, providing a reliable tool for clinical detection and epidemiological surveys of LSDV. This method offers significant potential for the prevention and control of LSD outbreaks.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"45"},"PeriodicalIF":3.5,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12139313/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144224183","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Dexamethasone-boosted mesenchymal stem cell secretome: insight into hepatic protection. 地塞米松促进间充质干细胞分泌组:洞察肝脏保护。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-06-04 DOI: 10.1186/s12896-025-00980-8
Eiman M Adly, Thoria Diab, Mohamed Hessien
{"title":"Dexamethasone-boosted mesenchymal stem cell secretome: insight into hepatic protection.","authors":"Eiman M Adly, Thoria Diab, Mohamed Hessien","doi":"10.1186/s12896-025-00980-8","DOIUrl":"10.1186/s12896-025-00980-8","url":null,"abstract":"<p><p>As the independent anti-inflammatory effects of mesenchymal stem cells (MSCs) and glucocorticoids are well documented, it is hypothesized that the conditioned media derived from dexamethasone (DEXA)-treated MSCs may exhibit a potent therapeutic effect. To explore this, bone marrow-derived MSCs were transiently maintained in DEXA-containing media, where cell viability, phenotype, and osteogenic differentiation were assessed. Furthermore, the MSC-conditioned media (MSC-CN) was utilized to inhibit the proliferation of hepatoma cells and treat drug-induced acute liver failure (ALF) in mice. We found that low doses of DEXA (≤100 nM) maintained MSC viability and their typical mesenchymal phenotype. Conversely, 1000 nM decreased the expression of the mesenchymal markers (CD105 and CD90), triggered osteogenic differentiation as evidenced by the modulation of osteogenesis-related genes (alkaline phosphatase, osteopontin, and Runt-related transcription factor 2), and increased the intracellular calcium, assessed by Alizarin Red S staining. Moreover, MSC-DEXA-S restricted colony formation, cell migration, and glucose consumption in hepatoma cells. In parallel, MSC-DEXA-S protected mice against acetaminophen-induced ALF, where both liver functions, oxidative stress (Nrf-2, SOD1, GSH, and MDA), angiogenic (VEGF), and inflammatory (TNF-α) markers were improved. Also, MSC-DEXA-S resolved liver necrosis one week after transfusion. These data suggest that pretreatment of MSCs with low doses of dexamethasone maintains their stemness and enhances their paracrine therapeutic effect against hepatic diseases.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"44"},"PeriodicalIF":3.5,"publicationDate":"2025-06-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12139281/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144224182","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Protocol optimization for callus induction and shoot regeneration of Ethiopian rice varieties (Oryza sativa L.). 埃塞俄比亚水稻品种愈伤组织诱导和茎部再生的方案优化。
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-05-24 DOI: 10.1186/s12896-025-00981-7
Teslim Yimam, Mereme Abide, Solomon Benor, Demisachew Guadie
{"title":"Protocol optimization for callus induction and shoot regeneration of Ethiopian rice varieties (Oryza sativa L.).","authors":"Teslim Yimam, Mereme Abide, Solomon Benor, Demisachew Guadie","doi":"10.1186/s12896-025-00981-7","DOIUrl":"10.1186/s12896-025-00981-7","url":null,"abstract":"<p><p>Rice is the primary food source for the majority of the global population. In African countries, rice is considered a critical cereal crop essential for food security. Rice consumption in Ethiopia has been increasing from year to year, and the demand for rice food is not fulfilled by domestic production, the Ethiopian government imports rice products from Asia. Therefore increasing the production of rice in terms of quantity and quality through modern technology is a recent priority and strategy in the country. This study was, therefore, designed to optimize the protocol for callus induction and shoot regeneration of two Ethiopian rice (X-Jigna and Shaga) varieties. Three media types (LS, N6, and MS) supplemented with different concentrations of 2, 4-D were used for callus induction. The maximum callus induction was observed on MS-media supplemented with 2.5 mg/L for X-Jigna and Shaga varieties, respectively. The lowest callus induction frequencies were recorded on N6-media supplemented with 1.5 mg/L 2, 4-D for both varieties. The highest shoot regeneration for X-Jigna (80.67%) and Shaga (72%) was recorded on MS-media containing a combination of kinetin (2 mg/L) and Naphtalin Acetic Acid (0.2 mg/L). The survival rate of acclimatized plantlets was 100% and 66.6% for X-Jigna and Shaga, respectively. In conclusion, this study is distinctive as, to our knowledge, no other research has addressed the optimization of protocols for the Ethiopian rice varieties, serving as a foundational element for future investigations into rice transformation and breeding in Ethiopia.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"43"},"PeriodicalIF":3.5,"publicationDate":"2025-05-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12103809/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144141423","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Assembly of ascovirus HvAV-3h long DNA fragment using the Transformation-Associated Recombination (TAR) approach in yeast cells. 利用转化相关重组(TAR)方法在酵母细胞中组装ascov -3h长DNA片段
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-05-22 DOI: 10.1186/s12896-025-00964-8
Heba A H Zaghloul, Zhengkun Xiao, Hengrui Hu, Guo-Hua Huang
{"title":"Assembly of ascovirus HvAV-3h long DNA fragment using the Transformation-Associated Recombination (TAR) approach in yeast cells.","authors":"Heba A H Zaghloul, Zhengkun Xiao, Hengrui Hu, Guo-Hua Huang","doi":"10.1186/s12896-025-00964-8","DOIUrl":"10.1186/s12896-025-00964-8","url":null,"abstract":"<p><strong>Background: </strong>Synthetic biology is a young but rapidly growing field that allows for assembling long DNA fragments, including complete chromosomes. A key approach for long-DNA assembly is the Transformation Associated Recombination (TAR), which relies on efficient homologous recombination in yeast cells. Recent reports indicate that the TAR method efficiently assembles some human and animal viruses characterized by their large DNA genome size. The application of the TAR method to synthesize long DNA fragments derived from insect viruses is scarce. Therefore, this study aimed to explore the TAR approach for the construction of a long DNA fragment (>44.6 Kb) from the insecticidal Heliothesis virescens ascovirus 3h (HvAV-3h) dsDNA genome to assess the suitability of this approach in genome-wide engineering studies in this family of viruses.</p><p><strong>Results: </strong>The long DNA fragment assembly process involved three stages: first, we amplified 15 segments of about 2.9-3.2 Kb each via PCR. Next, we recombined these segments through three parallel TAR cycles, producing medium-sized fragments of about 15 Kb. Finally, we assembled these fragments in a single TAR cycle to form a long DNA fragment of about 44.6 kb. We identified some positive clones by colony PCR or restriction digestion pattern. To assess the quality of the assembled DNA fragment, we conducted next-generation sequencing (NGS). A comparative analysis of Sanger sequencing for medium-sized fragments and NGS data from the synthesized long-DNA fragment demonstrated a nearly matched mutation profile, suggesting that the identified mutations and deletions were present at initial synthesis. Both datasets aligned with the reference HvAV-3h strain, revealing three specific nucleotide mutations and three unique mutation regions.</p><p><strong>Conclusions: </strong>Overall, the in vivo TAR assembly method efficiently assembled a long DNA fragment derived from the ascovirus genome as a template. The process is cost-effective and can be scaled up to synthesize the entire genome for gene functional studies.</p>","PeriodicalId":8905,"journal":{"name":"BMC Biotechnology","volume":"25 1","pages":"42"},"PeriodicalIF":3.5,"publicationDate":"2025-05-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12100921/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144126506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Bio-induced overproduction of heterocycloanthracin-like bacteriocin in Lysinibacillus macroides by Aspergillus austroafricanus: optimization of medium conditions and evaluation of potential applications. austroafricanus Aspergillus austroafricanus生物诱导的巨生赖氨酸芽胞杆菌过量产生异环蒽醌样细菌素:培养基条件优化及潜在应用评价
IF 3.5 3区 生物学
BMC Biotechnology Pub Date : 2025-05-22 DOI: 10.1186/s12896-025-00979-1
Philomena Edet, Maurice Ekpenyong, Atim Asitok, David Ubi, Cecilia Echa, Uwamere Edeghor, Sylvester Antai
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