LWT-FOOD SCIENCE AND TECHNOLOGY
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2024
·
IF 6
Junyan Liu,Zhufang Xiang,Tengyi Huang,Zhenbo Xu,Gongliang Liu,Chunyun Qu,Thanapop Soteyome,Yuting Luo,Qin Ma,Lei Yuan,Wei Hong,Xuejie Li,Yanrui Ye
Vibrio parahaemolyticus is a predominate cause of microbiological food safety. This study aimed to develop a polymerase spiral reaction (PSR) based assay for rapid and visual detection on V. parahaemolyticus . Firstly, the specificity and conservativity of species-specific target tlh gene were determined. Secondly, targeting tlh and virulence factors tdh and trh , PSR assay was developed and optimized using constructed positive plasmid controls and standard strain. Visualized result determination using fluorescent dye was applied to avoid false positive issues. Thirdly, 44 strains were used for specificity evaluation. Fourthly, pure and mixed species artificial contamination systems were included to test its applicability. In addition, a propidium monoazide (PMA)-PSR assay was set up to determine viable but non-culturable (VBNC) cells. Target tlh gene acquired high specificity and PSR assay was optimized to carry out at 65ᵒC for 1 h. Detection limit of PSR assay in purified genomic DNA, plasmid and artificially contaminated food samples were 5.31–53.1 pg/μL, 100 copies, and 10 3 CFU/mL, respectively, demonstrating the sensitivity as 100-fold higher than regular PCR. With PMA-PSR assay, VBNC cells were accurately determined without the disruption of dead cells and food compositions. Thus, PSR is applicable in detection of V. parahaemolyticus viable cells and virulence factors.
研究领域:
食品安全
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分子生物学
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微生物学
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DOI
高影响因子
Nature Water
·
2024
·Vol. 2
·期 6
·553-565
·
IF 0
Li Ruilong,An Xin-Li,Wang Yijin,Yang Zhugen,Su Jian-Qiang,Cooper Jonathan,Zhu Yong-Guan
Microplastics provide a unique niche for viruses, promoting viral interactions with hosts and accelerating the rapid ‘horizontal’ spread of antibiotic resistance genes (ARGs). Currently, however, there is a lack of knowledge concerning the main drivers for viral distribution on microplastics and on the resulting patterns of viral biogeographic distributions and the spread of the associated ARGs. Here we performed metagenomic and virus enrichment-based viromic sequencings on both polyethylene and polypropylene microplastics along a river. Experimental results show that Proteobacteria, Firmicutes, Actinobacteria and Cyanobacteria were the potential hosts of viruses on microplastics, but only approximately 4.1% of viral variations were associated with a bacterial community. Notably, two shared ARGs and six metal resistance genes were identified in both viral and their host bacterial genomes, indicating the occurrence of horizontal gene transfer between viruses and bacteria. Furthermore, microplastics introduce more distinctive elements to viral ecology, fostering viral diversification and virus–host linkage while refraining from an escalated level of horizontal gene transfer of ARGs in contrast to natural matrixes. Our study provides comprehensive profiles of viral communities, virus-related ARGs and their driving factors on microplastics, highlighting how these anthropogenic niches provide unique interfaces that comprise highly defined viral ecological features in the environment.
研究领域:
环境科学
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微生物学
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基因组学
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DOI
Nature Plants
·
2021
·Vol. 7
·期 8
·1026-1036
·
IF 15.793
Xiong Xingyao,Gou Junbo,Liao Qinggang,Li Yanlin,Zhou Qian,Bi Guiqi,Li Chong,Du Ran,Wang Xiaotong,Sun Tianshu,Guo Lvjun,Liang Haifei,Lu Pengjun,Wu Yaoyao,Zhang Zhonghua,Ro Dae-Kyun,Shang Yi,Huang Sanwen,Yan Jianbin
The ancient gymnosperm genus Taxus is the exclusive source of the anticancer drug paclitaxel, yet no reference genome sequences are available for comprehensively elucidating the paclitaxel biosynthesis pathway. We have completed a chromosome-level genome of Taxus chinensis var. mairei with a total length of 10.23 gigabases. Taxus shared an ancestral whole-genome duplication with the coniferophyte lineage and underwent distinct transposon evolution. We discovered a unique physical and functional grouping of CYP725A s (cytochrome P450) in the Taxus genome for paclitaxel biosynthesis. We also identified a gene cluster for taxadiene biosynthesis, which was formed mainly by gene duplications. This study will facilitate the elucidation of paclitaxel biosynthesis and unleash the biotechnological potential of Taxus .
研究领域:
植物生物学
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基因组学
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生物化学
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药理学
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产品关联
DOI
PubMed
高影响因子
ECOTOXICOLOGY AND ENVIRONMENTAL SAFETY
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2025
·
IF 6.1
Biyu Wu,Zirui Cheng,Xiang Li,Minxing Liang,Xue Wang,Duan Pi,Jiayi Liu,Huiling Li,Jun Zhao,Junjie Wang,Fang Liang,Xuegeng Wang
As a major substitute for the bisphenol A (BPA), the use of the bisphenol F (BPF) has increased dramatically in recent years. Growing evidence suggest that BPF shares numerous toxicological properties with BPA, raising the concerns about its potential impact on the health of organisms. However, the developmental toxicity of BPF remains poorly understood. In this study, we conducted a 5-day BPF exposure experiment on zebrafish ( Danio rerio ) from blastula stage at concentrations of 2, 20, 200, and 2000 µg/L. Our results demonstrated a significant increase in hatching rates across all treatment groups at 2 days post-fertilization (dpf). The esr1 was significantly upregulated at 2000 µg/L by 5 dpf, while no significant change was observed in ar . The frequency of operculum loss significantly increased at exposure concentrations of 20, 200, and 2000 µg/L, and a notable increase in notochord loss was observed at 2000 µg/L. To explore the underlying mechanisms, transcriptomic analysis was performed to identify differentially expressed genes (DEGs). GO and KEGG pathway enrichment analysis revealed that the toxic effects of BPF were closely associated with osteoclast differentiation, the FoxO signaling pathway, and the MAPK signaling pathway. These pathways influenced critical biological processes, including response to stimuli, animal organ morphogenesis, detoxification, and biomineralization. This study provides evidence that BPF exposure at environmentally relevant concentrations (2 µg/L) is harmful to hatching, concentrations above 20 µg/L exhibit estrogenic-disrupting activity and exert toxicological effects on the development of the head skeleton in zebrafish. These effects are particularly linked to disruptions in osteoclast differentiation.
研究领域:
环境毒理学
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分子生物学
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产品关联
DOI
PubMed
高影响因子
Scientific Reports
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2014
·Vol. 4
·期 1
·1-8
·
IF 5.078
Zhou Dinggang,Guo Jinlong,Xu Liping,Gao Shiwu,Lin Qingliang,Wu Qibin,Wu Luguang,Que Youxiong
To meet the demand for detection of foreign genes in genetically modified (GM) sugarcane necessary for regulation of gene technology, an efficient method with high specificity and rapidity was developed for the cry1Ac gene, based on loop-mediated isothermal amplification (LAMP). A set of four primers was designed using the sequence of cry1Ac along with optimized reaction conditions: 5.25 mM of Mg 2+ , 4:1 ratio of inner primer to outer primer, 2.0 U of Bst DNA polymerase in a reaction volume of 25.0 μL. Three post-LAMP detection methods (precipitation, calcein (0.60 mM) with Mn 2+ (0.05 mM) complex and SYBR Green I visualization), were shown to be effective. The sensitivity of the LAMP method was tenfold higher than that of conventional PCR when using templates of the recombinant cry1Ac plasmid or genomic DNA from cry1Ac transgenic sugarcane plants. More importantly, this system allowed detection of the foreign gene on-site when screening GM sugarcane without complex and expensive instruments, using the naked eye. This method can not only provide technological support for detection of cry1Ac , but can also further facilitate the use of this detection technique for other transgenes in GM sugarcane.
研究领域:
分子生物学
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植物生物技术
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基因工程
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产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2021
·
IF 4.821
Zhengzheng Wang,Guoyang Xie,Guanhua Chen,Xingcai Gao,Jiaxin Li,Zongliang Xie,Hengyi Xu
Due to the massive use of antibiotics, clinical highly pathogenic bacteria methicillin-resistant Staphylococcus aureus (MRSA) have emerged. Given its non-uniform drug resistance, it brings numerous difficulties for clinical diagnostic tests. This study aimed to establish a fast and accurate method to detect MRSA in hospital samples. In order to shorten the detection time and eliminate the influence of matrix on the detection results, so as to improve the detection sensitivity. In this study, magnetic separation (MS) technology was used for pre-treatment of samples, and triplex PCR was established using mecA , nuc and flic genes to realize the detection of MRSA. The results showed that the lowest detection limit of MRSA in blood samples and cerebrospinal fluid could reach 10 2 CFU/mL and 10 3 CFU/mL. Therefore, the MS technology combined with triplex PCR assay established in this study is expected to provide a strong basis for the detection of MRSA in hospital samples.
研究领域:
医学检验
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微生物学
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临床诊断
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产品关联
DOI
高影响因子
BioNanoScience
·
2025
·Vol. 15
·期 2
·1-10
·
IF 3.2
Cai He-Qing,Han Lu,Xue Xin-Yu,Zhang Kou,Wang Tian-Hao,Pan Fu-Hong,Sun Zhi-Cheng,Liu Ru-Ping,Hu Kun,Wei Yen
Gold nanocages (AuNCs) exhibit tunable local surface plasmon resonance (LSPR) properties and exceptional drug loading and releasing capabilities, attributes stemming from their hollow interiors and ultra-thin porous walls. These features underscore their promising applications in the biomedical field. In this study, we fabricated AuNCs with LSPR peaks at 810 nm via chemical etching of Ag nanocube templates with sharp edges using a precisely controlled volume of chloroauric acid. To impart gene-loading functionality, we chemically anchored thiol-modified polyethyleneimine (PEI) onto the surfaces of AuNCs through robust Au–S bonds, generating a novel gene carrier, AuNCs-PEI. Subsequently, we evaluated its photothermal efficacy using a femtosecond laser and assessed cytotoxicity via MTT assay. The binding affinity of AuNCs-PEI for EGFP plasmids was characterized by gel retardation assays. Furthermore, the gene delivery efficiency of AuNCs-PEI was examined in 293 T cells by quantifying green fluorescence protein signals using inverted fluorescence microscopy. These findings demonstrate that engineered AuNCs-PEI not only facilitates gene loading for therapeutic applications but also enables photothermal therapy (PTT).
研究领域:
生物医学工程
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纳米医学
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基因治疗
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产品关联
DOI
高影响因子
PLANT CELL REPORTS
·
2010
·Vol. 29
·期 8
·887-894
·
IF 2.301
Wang Cui-Ting,Liu Hua,Gao Xiao-Shu,Zhang Hong-Xia
A number of genes that function in the terpenoid indole alkaloids (TIAs) biosynthesis pathway have been identified in Catharanthus roseus . Except for the geraniol 10-hydroxylase (G10H) gene, which encodes a cytochrome P450 monooxygenase, several of these genes are up-regulated by ORCA3, a jasmonate-responsive APETALA2 (AP2)-domain transcript factor. In this study, the G10H gene was transformed independently, or co-transformed with ORCA3 into C. roseus , using Agrobacterium rhizogenes MSU440. Hairy root clones expressing the G10H gene alone, or both the G10H and ORCA3 genes, were obtained. Alkaloid accumulation level analyses showed that all transgenic clones accumulated more catharanthine, with the highest accumulation level in the transgenic clone OG12 (6.5-fold higher than that of the non-expression clone). Following treatment with ABA, accumulation of catharanthine reached 1.96 mg/g DW in the transgenic clone OG12. The expression levels of TIAs biosynthesis genes in transgenic and non-transgenic clones were also investigated.
研究领域:
植物学
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代谢工程
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药用植物研究
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DOI
PubMed
高影响因子
JOURNAL OF ASIA-PACIFIC ENTOMOLOGY
·
2024
·
IF 1.1
Qing-Hua Xiao,Wu-Ye Li,Jin Zhang,Jia-Min Yu,Dong-Yang Liu,Jiang-Nan Peng,Mao-Ye Li,Su Liu
In insects, the transcription factor cap ‘n’ collar isoform C (CncC) plays a critical role in the regulation of multiple genes involved in insecticide detoxification. Knockdown of CncC genes leads to increased susceptibility to different types of insecticides in several insect species. However, the CncC gene has not yet been fully characterized in the black cutworm Agrotis ipsilon , a notorious insect pest that causes severe damage to field crops. In this study, the CncC gene (designated AiCncC ) was identified from A. ipsilon . Exposure to a median lethal concentration (LC 50 ) of chlorantraniliprole (CAP) or phoxim (PHO) strongly increased the expression of AiCncC . Silencing of AiCncC by RNA interference significantly increased the susceptibility of A. ipsilon larvae to both insecticides. Moreover, CncC/Maf binding sites were predicted in the putative promoters of two glutathione S -transferase (GST) genes ( AiGSTe1 and AiGSTu1 ) involved in the detoxification of CAP and PHO. The transcription levels of AiGSTe1 and AiGSTu1 were dramatically decreased by silencing AiCncC . These findings indicate that AiCncC is associated with CAP and PHO susceptibility through the regulation of GST genes.
研究领域:
昆虫学
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分子生物学
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害虫防治
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产品关联
DOI
高影响因子
ENVIRONMENTAL SCIENCE AND POLLUTION RESEARCH
·
2015
·Vol. 22
·期 21
·16914-16926
·
IF 2.828
Wang Sheng,Bai Naling,Wang Bing,Feng Zhuo,Hutchins William C.,Yang Ching-Hong,Zhao Yuhua
Commonly used flame retardants, such as polybrominated diphenyl ethers, are extremely persistent in the environment, causing serious environmental risks. Certain strains of bacteria are able to degrade several low brominated congeners of PBDEs aerobically. However, the aerobic degradation pathway is not yet well understood, particularly at the genetic level. In this study, we isolated Cupriavidus sp. WS from the environment that could degrade diphenyl ether (DE), 4-bromodiphenyl ether, and 4,4′-bromodiphenyl ether. DE was completely degraded in 6 days without any detectable end-product. Using transposon mutagenesis, several DE degradation-deficient mutants were obtained. Knocking out bphA1 , bphA2 , and bphA3 eliminated the ability of the Cupriavidus sp. WS bacterium to degrade DE, indicating that the bph genes play a crucial role in DE degradation by this strain. The specific roles of bphA , bphB , and bphC were identified by systematically expressing these genes in Escherichia coli . The dihydrodiol product of BphA was dehydrogenated into 2,3-dihydroxydiphenyl ether by BphB. 2,3-Dihydroxydiphenyl ether was then decomposed into phenol and 2-pyrone-6-carboxylic acid by BphC. Thus, BphA, BphB, and BphC act sequentially in the aerobic degradation of DE, 4-bromodiphenyl ether, and 4,4′-dibromodiphenyl ether by the Cupriavidus sp. WS bacterium.
研究领域:
环境微生物学
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分子生物学
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污染治理
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DOI
PubMed
高影响因子
JOURNAL OF APPLIED MICROBIOLOGY
·
2020
·Vol. 129
·期 5
·1410-1418
·
IF 3.066
Li Z.,Liu Y.,Li Y.,Lin F.,Wu L.
AimsRed yeast rice (RYR), produced by inoculating Monascus strains to steamed rice, contains many kinds of physiologically bioactive compounds, among which monacolin K can be used as an antihypercholesterolaemic agent. However, RYR can be polluted by the mycotoxin citrinin, which has nephrotoxic and hepatotoxic activities. To avoid the risk of citrinin contamination in Monascus fermented products, it is important to screen for Monascus strains that produce no or low citrinin.Methods and ResultsFive autochthonous Monascus strains with high‐yield monacolin K and undetectable citrinin were obtained using high‐performance liquid chromatography (HPLC). All five strains were identified as Monascus ruber based on Genealogical Concordance Phylogenetic Species Recognition criteria. Polymerase chain reaction revealed that citrinin polyketide synthase (pksCT) gene was found in these strains, but transcriptional regulator (ctnA) was not found.ConclusionsFive strains are potential strains for producing high‐quality RYR. The distribution of the pksCT gene was not restricted to Monascus purpureus and Monascus sanguineus, and M. ruber strains were diverse in pksCT and ctnA genes.Significance and Impact of the StudyThe integration of citrinin HPLC analysis and pksCT and ctnA genes amplification could provide a complementary approach in valuable Monascus strains screening.
研究领域:
微生物学
·
食品科学
·
分子生物学
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产品关联
DOI
PubMed
高影响因子
JOURNAL OF MOLECULAR CATALYSIS B-ENZYMATIC
·
2013
·
IF 2.823
Xiaolin Pei,Lirong Yang,Gang Xu,Qiuyan Wang,Jianping Wu
Microbial nitrile hydratases (NHases) are important industrial catalysts to produce valuable amides. However, only some NHase genes have been reported and studied at the molecular level. In this study, we developed a genome mining method to discover Fe-type NHases from GenBank. The putative NHase gene from Pseudomonas putida F1 was cloned and functionally expressed in Escherichia coli BL21 (DE3) by assisting of a putative activator gene adjacent to β-subunit region. Three recombinant plasmids containing NHase gene or the activator gene were designed and constructed. Maximal enzyme activity was obtained when the structural and activator genes were transcribed as one unit in plasmid pCDFDuet-1 at 18 °C. However, the expressed product did not show any NHase activity when the downstream activator gene was ignored, and the product completely existed in insoluble inclusion body. The activator gene might be involved in protein folding of the α- and β-subunits of NHase. In addition, the Fe-type NHase exhibited broad substrate specificity. The enzyme can efficiently hydrate aromatic nitriles, such as 3-cyanopyridine, 4-cyanopyridine, and benzonitrile, asides from aliphatic nitriles preferentially. Therefore, the recombinant NHase shows potential applications in some amides preparation.
研究领域:
生物化学
·
微生物学
·
工业酶学
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产品关联
DOI
高影响因子
危险品化学品经营许可证（带存储）