BIOCHEMICAL ENGINEERING JOURNAL
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2026
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IF 3.6
Haobo Xing,Qingming Hou,Zhenhua Wang,Shanfei Zhang,Shangxia Song,Lulu Du,Chenghua Gao,Yunfei Li,Fubao Sun
Monensin is a polyether antibiotic produced by Streptomyces cinnamonensis . Due to its highly complex biosynthetic pathway, incomplete characterization of key biosynthetic genes has long hindered improvements in industrial-scale monensin production. Based on our previous transcriptomic analysis that key genes involved in monensin biosynthesis were initially identified in a high-yield S. cinnamonensis strain, this study focused on metabolic engineering of this strain through multi-gene integration strategy, targeting primary metabolism, precursor synthesis, ATP supply, stress resistance, and the monensin biosynthetic gene cluster. Results demonstrated that the co-expression of phosphofructokinase ( pfk ), citrate synthase ( cs ), methylmalonyl-CoA mutase subunit β ( mutB ), and methylmalonyl-CoA mutase ( mcm ) rendered the monensin titer from 12.6 g/L to 20.2 g/L, an approximately 60% increase compared with the original strain. Co-expression of the ATP synthase β subunit ( assβ ), the fungal thiol-coupled enzyme ( mac ), and the monensin biosynthetic enzyme ( monE ) allowed the monensin production from 12.6 g/L to 19.4 g/L, corresponding to an increase of above 50%. Furthermore, simultaneous co-expression of the seven genes using the novel pKHBT1 system increased the monensin titer to 26.2 g/L in a 5-L fermenter, more than double that of the original strain. These newly identified key metabolic genes were indeed associated closely with the monensin biosynthesis in S. cinnamonensis , which supplied a clue to metabolic engineering of the monensin production strain for efficient biosynthesis.
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高影响因子
CELLULAR SIGNALLING
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2023
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IF 4.8
Hao Zhou,Jiangbo Huang,Fang Wang
Background Studies are ongoing to examine the versatile functions of circular RNAs (circRNAs) in human diseases. This research investigates the effects of hsa_circ_0000644 (circ_644) and its related molecules on the malignant behavior of bladder cancer (BCa) cells. Methods Abundant bioinformatics analyses were performed to screen the key circRNA and its related molecules in BCa. Tumor tissues and the para-tumorous tissues were collected from 58 patients with BCa. Expression of RUNX family transcription factor 3 (RUNX3), circ_644, microRNA-143-3p (miR-143-3p), and musashi RNA binding protein 2 (MSI2) in BCa tissues or cells was determined. Molecular interactions were confirmed by chromatin immunoprecipitation, RNA pull-down, and luciferase assays. Gain and loss-of function assays were performed using two BCa cell lines (T24 and HT1376). Results Circ_644 was highly expressed whereas RUNX3 , which could suppress circ_644 transcription, was lowly expressed in BCa tissues and cells. Upregulation of RUNX3 suppressed proliferation, colony formation, migration and invasion, and tumorigenicity of BCa cells and induced cell cycle arrest . However, the tumor-suppressive effects of RUNX3 were blocked by circ_644 upregulation. Circ_644 served as a sponge for miR-143-3p, and miR-143-3p bound to MSI2 mRNA. The rescue experiments showed that miR-143-3p inhibition or MSI2 overexpression restored the malignant behaviors of BCa cells induced by circ_644 knockdown or RUNX3 overexpression. Conclusion This study demonstrates that transcriptional activation of circ_644 upon RUNX3 downregulation drives the malignant development of BCa through the miR-143-3p/MSI2 axis. RUNX3 restoration or specific inhibition of circ_644 or MSI2 may help block BCa progression.
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高影响因子
RESEARCH IN VETERINARY SCIENCE
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2016
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IF 1.504
Yong Sun,Tingting Guo,Dawei Guo,Li Guo,Li Chen,Yu Zhang,Liping Wang
Chicken P-glycoprotein (chP-gp), encoded by Abcb1 , determines the bioavailability because of its effect on pharmacokinetics of various drugs. However, comprehensive studies on chP-gp are still limited. In this study, the chicken full-length cDNA was first successfully cloned and then stably expressed in MDCK cell line. The open reading frame of chicken Abcb1 consists of 3864 nucleotides, encoding for a 1287-amino acid protein. Sequence alignments analysis showed that chicken P-gp had high identities with the homologues of turkey (95%), human (72%), pig (72%), rat (71%) and cattle (68%). The efflux ratio of rhodamine123 (Rho123, a human P-gp substrate) in ch Abcb1 transfected MDCK cells was significantly higher than that in the wild type MDCK cell (6.24 vs 1.64, P < 0.05), suggesting a good transporting function of chicken P-gp overexpressed in the transfected cell. Importantly, MDCK-ch Abcb1 cells, unlike Caco-2 cells, exhibited biphasic saturation kinetics in transporting Rho123. In conclusion, an MDCK cell line stably expressing ch Abcb1 was successfully established, which could provide a new cell model to screen its substrates and inhibitors and study the drug-drug interaction medicated via chicken P-gp.
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高影响因子
MOLECULAR BIOLOGY REPORTS
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2025
·Vol. 52
·期 1
·1-10
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IF 2.8
Wang Zhengwei,Lin Jiani,Tan Guisen,Pan Tingzheng,Zhang Hongtian,Liu Jiangang
Background Nuclear factor IX (NFIX) promotes glioblastoma (GBM) development by inducing GBM cell migration and proliferation. However, the upstream regulatory mechanisms of NFIX in GBM remain unclear. Methods and results To investigate the functional role of miR-1270 in GBM, a stable miR-1270-overexpressing cell model was constructed using lentiviral transduction. The functional consequences were systematically evaluated using Transwell migration, colony formation, and CCK-8 assays, as well as western blotting and quantitative real-time polymerase chain reaction. To validate the therapeutic potential in vivo, we established orthotopic xenograft models in nude mice and longitudinally monitored tumor progression through non-invasive small animal imaging, enabling comprehensive assessment of the antitumor effects of miR-1270 within a physiological microenvironment. A decrease in microRNA-1270 (miR-1270) expression in human GBM samples was observed, which was negatively correlated with NFIX expression. The miR-1270 inhibited NFIX expression, a key driver of GBM cell proliferation and migration. Mechanistic analysis revealed that miR-1270 directly bound to the NFIX promoter region, suppressing its transcriptional activity. The reintroduction of NFIX counteracted the inhibitory effects of miR-1270 on GBM cell malignancy. Conclusions This study is the first to report the transcriptional inhibition of NFIX by miR-1270 in GBM cells. These findings suggest that targeting the miR-1270–NFIX axis may provide a promising therapeutic strategy for GBM.
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JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
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2025
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IF 6.2
Mengjun Yun,Yu Xiong,Zhuobing Wang,Lianjie Xie,Hanwen Ye,Xiaofang Yuan,Weiyi He,Binqing Chen,Zhanjun Lu,Wei Chen
Plants perceive proteins from insect-derived oral secretion (OS) and regulate the classical endogenous hormone jasmonic acid to resist insects, but the role of abscisic acid (ABA) in this process is poorly understood. In this study, we used the specialist herbivorous caterpillar Plutella xylostella and cruciferous plants as a model to investigate how the ABA hormone responds to the OS and its core peptide from the insect. Through proteomics and Western blotting analysis, glucosinolate sulfatase 1 (GSS1) was identified in OS. Yeast library screening revealed that GSS1 and its 28-amino-acid core peptide (GSS1-P1) interact with ABA biosynthetic enzyme ABA1. Arabidopsis overexpressing GSS1 and plants treated with synthetic GSS1-P1 showed elevated ABA levels. Transcriptome analysis and RT-qPCR confirmed that GSS1-P1 upregulates WRKY18 and ABA1 expression, modulating ABA production. Both GSS1-P1 application and optimal ABA concentrations enhanced plant resistance to herbivory. Our study shows that GSS1 and its peptide stimulate ABA production, boosting plant–insect resistance and highlighting ABA’s potential role in pest-stress response.
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Microbiology Spectrum
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2025
·Vol. 13
·期 10
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IF 3.8
Daoyan Tang,Minggui Wang,Dan Wang,Danni Yang,Yi Cai,Tao Luo,Jianqing He,Qinglan Wang
Efficient DNA delivery is essential for genetic manipulation of mycobacteria and for dissecting their physiology, pathogenesis, and drug resistance. Although electroporation enables transformation efficiencies exceeding 10⁵ CFU per µg DNA in Mycobacterium smegmatis and Mycobacterium tuberculosis, it remains highly inefficient in many nontuberculous mycobacteria (NTM), including Mycobacterium abscessus. Here, we discovered that NTM such as M. abscessus exhibit exceptional tolerance to ultra-high electric field strengths and that hypertonic preconditioning partially protects cells from electroporation-induced damage. Using ultra-high electric field strength (3 kV/mm) electroporation, we achieved dramatic improvements in plasmid transformation efficiency—up to 106-fold in M. abscessus, 83-fold in Mycobacterium marinum, and 37-fold in Mycobacterium kansasii—compared to standard conditions (1.25 kV/mm). Transformation efficiency was further influenced by the choice of selectable marker. Ultra-high field strength electroporation also markedly enhanced allelic exchange in M. abscessus expressing Che9c RecET recombinases, increasing the recovery of gene deletion mutants by over 1,000-fold relative to conventional electroporation. In parallel, oligonucleotide-mediated recombineering for targeted point mutations produced nearly 10,000-fold more mutants under ultra-high field conditions. Together, these findings establish ultra-high field electroporation as a robust, broadly applicable platform for genetic engineering of NTMs. This method substantially enhances transformation efficiency and enables construction of advanced genetic tools—including expression libraries and CRISPRi knockdown libraries—in species that have historically resisted genetic manipulation.
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高影响因子
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