The word “natto” originated from Japan, which was known as stinky bean, salt bean or silk bean in Chinese folk. Natto contains multiple functional components, including nattokinase, soybean isoflavone, γ-polyglutam...The word “natto” originated from Japan, which was known as stinky bean, salt bean or silk bean in Chinese folk. Natto contains multiple functional components, including nattokinase, soybean isoflavone, γ-polyglutamic acid, vitamin K2, biogenic amines, SOD, small-molecule polypeptide, which has good preventive and curative effects for many diseases, such as cardiovascular and cerebrovascular diseases, hyperlipidemia, hypertension, menopausal symptoms and hypoimmunity. Along with the aging population in the world and China, it is urgent to improve the quality of life. It is necessary to enhance the understanding of natto-related knowledge, expand propaganda power, accept natto consciously and eat natto frequently, which is good for our health and helps our country to build a healthy and harmonious society. This review will descript the development, components, functions and prospects of natto, which help us to offer theoretical basis for promoting natto industry.展开更多
Nuclear power is essential for sustainable energy infrastructure and economic development,necessitating materials for high-radiation environments that can facilitate visualization and observation.Conventional lead gla...Nuclear power is essential for sustainable energy infrastructure and economic development,necessitating materials for high-radiation environments that can facilitate visualization and observation.Conventional lead glass is inadequate for future requirements due to radiation-induced darkening,poor mechanical properties,and toxicity.Therefore,there is urgent to find new window materials that offer multi-ionization shielding(particularly against deep-penetrating gamma ray,γ,and neutron,n,radiations),desirable opto-mechanical properties,service stability against darkening,and non-toxicity.In this study,we report a family of transparent rare-earth pyrochlore ceramics La_(x)Gd_(2-x)Zr_(2)O_(7),offering unique chemo-physical properties that are ideal for robust radiation shielding windows.Remarkably,we demonstrated the capability of maintaining high transparency under heavy-dose exposure to 1000 kGy ^(60)Coγradiation.We observed the service stability against radiation darkening can be greatly enhanced with La-rich compositions,while Gd-rich compositions undergo shallow darkening that can be reversibly recovered under visible light.This behavior is attributed to mitigated oxygen migration from 48f to 8a in La-rich compositions,which have high pyrochlore phase stability and well-ordered atomic structures,and reversible oxygen migration between 48f and 8a in Gd-rich compositions,which remain active at room temperature.Our proposal and demonstration unlock ample opportunities in designing functional transparent ceramics as window materials for demanding applications in high-radiation environments.展开更多
Human endogenous retrovirus W family(HERV-W) envelope(env) has been reported to be related to several human diseases, including autoimmune disorders, and it could activate innate immunity.However, there are no reports...Human endogenous retrovirus W family(HERV-W) envelope(env) has been reported to be related to several human diseases, including autoimmune disorders, and it could activate innate immunity.However, there are no reports investigating whether human leukemia antigen(HLA)-A~*0201^+restriction is involved in the immune response caused by HERV-W env in neuropsychiatric diseases. In the present study, HERV-W env-derived epitopes presented by HLA-A~*0201 are described with the potential for use in adoptive immunotherapy. Five peptides displaying HLAA~*0201-binding motifs were predicted using SYFEPITHI and BIMAS, and synthesized. A CCK-8 assay showed peptides W, Q and T promoted lymphocyte proliferation. Stimulation of peripheral blood mononuclear cells from HLA-A~*0201^+ donors with each of these peptides induced peptidespecific CD8^+ T cells. High numbers of IFN-γ-secreting T cells were also detectable after several weekly stimulations with W, Q and T. Besides lysis of HERV-W env-loaded target cells, specific apoptosis was also observed. These data demonstrate that human T cells can be sensitized toward HERV-W env peptides(W, Q and T) and, moreover, pose a high killing potential toward HERV-W env-expressing U251 cells. In conclusion, peptides W Q and T, which are HERV-W env antigenic epitopes, have both antigenicity and immunogenicity, and can cause strong T cell immune responses. Our data strengthen the view that HERV-W env should be considered as an autoantigen that can induce autoimmunity in neuropsychiatric diseases, such as multiple sclerosis and schizophrenia. These data might provide an experimental foundation for a HERV-W env peptide vaccine and new insight into the treatment of neuropsychiatric diseases.展开更多
Electrochemical conversion with solid oxide electrolysis cells is a promising technology for CO2 utilization and simultaneously store renewable energy.In this work,Ce0.9M0.1O2-δ(CeM,M=Fe,Co,Ni)catalysts are infiltrat...Electrochemical conversion with solid oxide electrolysis cells is a promising technology for CO2 utilization and simultaneously store renewable energy.In this work,Ce0.9M0.1O2-δ(CeM,M=Fe,Co,Ni)catalysts are infiltrated into La0.6Sr0.4Cr0.5Fe0.5O3-δ-Gd0.2Ce0.8O2-δ(LSCr Fe-GDC)cathode to enhance the electrochemical performance for CO2 electrolysis.CeCo-LSCrFe-GDC cell obtains the best performance with a current density of 0.652 A cm^-2,followed by CeFe-LSCrFe-GDC and CeNi-LSCrFe-GDC cells with the value of 0.603 and 0.535 A cm^-2,respectively,about 2.44,2.26 and 2.01 times higher than that of the LSCrFe-GDC cell at1.5 V and 800℃.Electrochemical impedance spectra combined with distributions of relaxed times analysis shows that both CO2 adsorption process and the dissociation of CO2 at triple phase boundaries are accelerated by Ce M catalysts,while the latter is the key rate-determining step.展开更多
Human endogenous retrovirus W env(HERV-W env) plays a critical role in many neuropsychological diseases such as schizophrenia and multiple sclerosis(MS). These diseases are accompanied by immunological reactions in th...Human endogenous retrovirus W env(HERV-W env) plays a critical role in many neuropsychological diseases such as schizophrenia and multiple sclerosis(MS). These diseases are accompanied by immunological reactions in the central nervous system(CNS). Microglia are important immunocytes in brain inflammation that can produce a gasotransmitter – nitric oxide(NO). NO not only plays a role in the function of neuronal cells but also participates in the pathogenesis of various neuropsychological diseases. In this study, we reported increased NO production in CHME-5 microglia cells after they were transfected with HERV-W env. Moreover, HERV-W env increased the expression and function of human inducible nitric oxide synthase(hi NOS) and enhanced the promoter activity of hi NOS. Microglial migration was also enhanced. These data revealed that HERV-W env might contribute to increase NO production and microglial migration ability in neuropsychological disorders by regulating the expression of inducible NOS. Results from this study might lead to the identification of novel targets for the treatment of neuropsychological diseases, including neuroinflammatory diseases, stroke, and neurodegenerative diseases.展开更多
Abiotic stress such as high temperature at flowering is one of many conditions reducing yield of corn(Zea mays L.).Mixing corn cultivars with diverse functional traits increases within-crop diversity and provides a po...Abiotic stress such as high temperature at flowering is one of many conditions reducing yield of corn(Zea mays L.).Mixing corn cultivars with diverse functional traits increases within-crop diversity and provides a potential means of mitigating yield losses under stress conditions.We conducted a three-year field study to investigate the effects of cultivar mixtures on kernel setting rate,pollen sources,and yield.This study consisted of six treatments,including two high temperature-tolerant(HTT)monocrops of WK702 and DH701,two high temperature-sensitive(HTS)monocrops of DH605 and DH662,and two HTT–HTS mixtures of WK702-DH605 and DH701-DH662.The anthesis–silking interval(ASI)was 0.9–1.6 days shorter in mixtures than in monocrops.Kernel setting rate was increased in mixtures(86.4%–88.7%)compared with those in monocrops(74.7%–84.1%)as a result of synchrony and complementarity of pollination.Grain yields of the HTT–HTS mixtures increased by 13.3%–18.7%,equivalent to 1169 to1605 kg ha^(-1),in comparison with HTS corn monocrops.The results of SSR markers showed that crossfertilization percentage in corn cultivar mixtures ranged from 29.3%to 47.8%,partially explaining yield improvement.Land equivalent ratio(LER)was 1.12 for corn mixtures and the partial land equivalent ratio(e.g.,>0.5)showed the complementary benefits in corn mixtures.The results indicated that mixing corn cultivars with diverse flowering and drought-tolerance traits increased yields via pollination synchrony.展开更多
Due to its high efficiency, high precision and high flexibility, CNC lathe is widely used in the machinery manufacturing industry increasingly, and becomes one of CNC machine too[s that most widely used. However, to g...Due to its high efficiency, high precision and high flexibility, CNC lathe is widely used in the machinery manufacturing industry increasingly, and becomes one of CNC machine too[s that most widely used. However, to give full play to the role of CNC lathes, the key is programme, that is, preparing the reasonable and efficient processing procedures depending on the features and precision parts. This paper discussed the problems of programming and processing techniques of the CNC lathe parts.展开更多
Waterlogging stress significantly impairs plant growth and reduces crop yields.Spermidine(Spd),functioning as a second messenger,demonstrates positive effects on plant growth under waterlogging stress conditions.Howev...Waterlogging stress significantly impairs plant growth and reduces crop yields.Spermidine(Spd),functioning as a second messenger,demonstrates positive effects on plant growth under waterlogging stress conditions.However,the molecular mechanisms by which exogenous Spd application alleviates waterlogging stress remain unclear.This study employed physiological analysis and multi-omics approaches to investigate the effect of Spd application on waterlogging stress.The application of Spd enhanced the expression of genes related to light-harvesting complex(LHC),photosynthesis,and starch-related pathways,while inhibiting chlorophyll degradation and maintaining higher photosynthetic rates,thereby increasing biomass accumulation under waterlogging stress.The activation of genes associated with trehalose and Spd biosynthesis resulted in elevated accumulation of trehalose and endogenous Spd.The inhibition of 1-aminocyclopropane-1-carboxylic acid(ACC)oxidase(ACO)expression contributed to reduced ethylene emission,enhancing maize resistance to waterlogging.Following Spd application,auxin-related genes were up-regulated and indole acetic acid(IAA)content increased,promoting cell elongation in maize and maintaining normal growth under waterlogging stress.Additionally,the upregulation of lipid-related genes led to increased lipid content,protecting cell membranes under waterlogging conditions.These molecular and physiological modifications collectively enhanced resistance to waterlogging stress.These findings advance our understanding of Spd's regulatory roles in mitigating waterlogging damage and provide valuable insights for breeding waterlogging-tolerant maize varieties.展开更多
Accurately counting dense objects in complex and diverse backgrounds is a significant challenge in computer vision,with applications ranging from crowd counting to various other object counting tasks.To address this,w...Accurately counting dense objects in complex and diverse backgrounds is a significant challenge in computer vision,with applications ranging from crowd counting to various other object counting tasks.To address this,we propose HUANNet(High-Resolution Unified Attention Network),a convolutional neural network designed to capture both local features and rich semantic information through a high-resolution representation learning framework,while optimizing computational distribution across parallel branches.HUANNet introduces three core modules:the High-Resolution Attention Module(HRAM),which enhances feature extraction by optimizing multiresolution feature fusion;the Unified Multi-Scale Attention Module(UMAM),which integrates spatial,channel,and convolutional kernel information through an attention mechanism applied across multiple levels of the network;and the Grid-Assisted Point Matching Module(GPMM),which stabilizes and improves point-to-point matching by leveraging grid-based mechanisms.Extensive experiments show that HUANNet achieves competitive results on the ShanghaiTech Part A/B crowd counting datasets and sets new state-of-the-art performance on dense object counting datasets such as CARPK and XRAY-IECCD,demonstrating the effectiveness and versatility of HUANNet.展开更多
Bacillus subtilis plays an important role in fundamental and applied research,and it has been widely used as a cell factory for the production of enzymes,antimicrobial materials,and chemicals for agriculture,medicine,...Bacillus subtilis plays an important role in fundamental and applied research,and it has been widely used as a cell factory for the production of enzymes,antimicrobial materials,and chemicals for agriculture,medicine,and industry.However,genetic manipulation tools for B.subtilis have low efficiency.In this work,our goal was to develop a simple recombineering system for B.subtilis.We showed that genome editing can be achieved in B.subtiliis 1A751 through co-expression of YqaJ/YqaK,a native phage recombinase pair found in B.sub-tilis 168,and the competence master regulator ComK using a double-stranded DNA substrate with short ho-mology arms(100 bp)and a phosphorothioate modification at the 5′-end.Efficient gene knockouts and large DNA insertions were achieved using this new recombineering system in B.subtilis 1A751.As far as we know,this is the first recombineering system using the native phage recombinase pair YqaJ/YqaK in B.subtilis.In conclusion,this new recombineering system provides a simple and fast tool for genetic manipulation of B.sub-tilis,and it will promote studies of genome function,construction of production strains,and genome mining in B.subtilis.展开更多
文摘The word “natto” originated from Japan, which was known as stinky bean, salt bean or silk bean in Chinese folk. Natto contains multiple functional components, including nattokinase, soybean isoflavone, γ-polyglutamic acid, vitamin K2, biogenic amines, SOD, small-molecule polypeptide, which has good preventive and curative effects for many diseases, such as cardiovascular and cerebrovascular diseases, hyperlipidemia, hypertension, menopausal symptoms and hypoimmunity. Along with the aging population in the world and China, it is urgent to improve the quality of life. It is necessary to enhance the understanding of natto-related knowledge, expand propaganda power, accept natto consciously and eat natto frequently, which is good for our health and helps our country to build a healthy and harmonious society. This review will descript the development, components, functions and prospects of natto, which help us to offer theoretical basis for promoting natto industry.
基金supported by National Key Research and Development Program of China,China(grant no.2023YFB3812000)National Natural Science Foundation of China,China(U21A20441)Science and Technology Project of Sichuan Province,China(2023ZYD0137).
文摘Nuclear power is essential for sustainable energy infrastructure and economic development,necessitating materials for high-radiation environments that can facilitate visualization and observation.Conventional lead glass is inadequate for future requirements due to radiation-induced darkening,poor mechanical properties,and toxicity.Therefore,there is urgent to find new window materials that offer multi-ionization shielding(particularly against deep-penetrating gamma ray,γ,and neutron,n,radiations),desirable opto-mechanical properties,service stability against darkening,and non-toxicity.In this study,we report a family of transparent rare-earth pyrochlore ceramics La_(x)Gd_(2-x)Zr_(2)O_(7),offering unique chemo-physical properties that are ideal for robust radiation shielding windows.Remarkably,we demonstrated the capability of maintaining high transparency under heavy-dose exposure to 1000 kGy ^(60)Coγradiation.We observed the service stability against radiation darkening can be greatly enhanced with La-rich compositions,while Gd-rich compositions undergo shallow darkening that can be reversibly recovered under visible light.This behavior is attributed to mitigated oxygen migration from 48f to 8a in La-rich compositions,which have high pyrochlore phase stability and well-ordered atomic structures,and reversible oxygen migration between 48f and 8a in Gd-rich compositions,which remain active at room temperature.Our proposal and demonstration unlock ample opportunities in designing functional transparent ceramics as window materials for demanding applications in high-radiation environments.
基金supported by grants from the National Natural Sciences Foundation of China(no.31470264,no.81271820,no.30870789 and no.30300117)the Key Program of the Natural Science Foundation of Hubei Province of China(no.2014CFA078)+1 种基金the Stanley Foundation from the Stanley Medical Research Institute(SMRI),USA(no.06R-1366),for Dr.F Zhuthe Scientific Innovation Team Project of Hubei Province of China(no.2015CFA009)
文摘Human endogenous retrovirus W family(HERV-W) envelope(env) has been reported to be related to several human diseases, including autoimmune disorders, and it could activate innate immunity.However, there are no reports investigating whether human leukemia antigen(HLA)-A~*0201^+restriction is involved in the immune response caused by HERV-W env in neuropsychiatric diseases. In the present study, HERV-W env-derived epitopes presented by HLA-A~*0201 are described with the potential for use in adoptive immunotherapy. Five peptides displaying HLAA~*0201-binding motifs were predicted using SYFEPITHI and BIMAS, and synthesized. A CCK-8 assay showed peptides W, Q and T promoted lymphocyte proliferation. Stimulation of peripheral blood mononuclear cells from HLA-A~*0201^+ donors with each of these peptides induced peptidespecific CD8^+ T cells. High numbers of IFN-γ-secreting T cells were also detectable after several weekly stimulations with W, Q and T. Besides lysis of HERV-W env-loaded target cells, specific apoptosis was also observed. These data demonstrate that human T cells can be sensitized toward HERV-W env peptides(W, Q and T) and, moreover, pose a high killing potential toward HERV-W env-expressing U251 cells. In conclusion, peptides W Q and T, which are HERV-W env antigenic epitopes, have both antigenicity and immunogenicity, and can cause strong T cell immune responses. Our data strengthen the view that HERV-W env should be considered as an autoantigen that can induce autoimmunity in neuropsychiatric diseases, such as multiple sclerosis and schizophrenia. These data might provide an experimental foundation for a HERV-W env peptide vaccine and new insight into the treatment of neuropsychiatric diseases.
基金financially supported by the National Natural Science Foundation of China (Nos. 91534128, 21506208 and 21476230)the Ministry of Science and Technology of China (Grants 2016YFE0118300)the DNL Cooperation Fund, CAS (DNL180306)
文摘Electrochemical conversion with solid oxide electrolysis cells is a promising technology for CO2 utilization and simultaneously store renewable energy.In this work,Ce0.9M0.1O2-δ(CeM,M=Fe,Co,Ni)catalysts are infiltrated into La0.6Sr0.4Cr0.5Fe0.5O3-δ-Gd0.2Ce0.8O2-δ(LSCr Fe-GDC)cathode to enhance the electrochemical performance for CO2 electrolysis.CeCo-LSCrFe-GDC cell obtains the best performance with a current density of 0.652 A cm^-2,followed by CeFe-LSCrFe-GDC and CeNi-LSCrFe-GDC cells with the value of 0.603 and 0.535 A cm^-2,respectively,about 2.44,2.26 and 2.01 times higher than that of the LSCrFe-GDC cell at1.5 V and 800℃.Electrochemical impedance spectra combined with distributions of relaxed times analysis shows that both CO2 adsorption process and the dissociation of CO2 at triple phase boundaries are accelerated by Ce M catalysts,while the latter is the key rate-determining step.
基金supported by grants from the National Natural Sciences Foundation of China(No.31470264,No.81271820,No.30870789,and No.30300117)the Key Program of Natural Science Foundation of Hubei Province of China(No.2014CFA078)+1 种基金the Stanley Foundation from the Stanley Medical Research Institute(SMRI),USA(No.06R-1366),to Dr.Fan Zhuthe Scientific Innovation Team Project of Hubei Province of China(No.2015CFA009)
文摘Human endogenous retrovirus W env(HERV-W env) plays a critical role in many neuropsychological diseases such as schizophrenia and multiple sclerosis(MS). These diseases are accompanied by immunological reactions in the central nervous system(CNS). Microglia are important immunocytes in brain inflammation that can produce a gasotransmitter – nitric oxide(NO). NO not only plays a role in the function of neuronal cells but also participates in the pathogenesis of various neuropsychological diseases. In this study, we reported increased NO production in CHME-5 microglia cells after they were transfected with HERV-W env. Moreover, HERV-W env increased the expression and function of human inducible nitric oxide synthase(hi NOS) and enhanced the promoter activity of hi NOS. Microglial migration was also enhanced. These data revealed that HERV-W env might contribute to increase NO production and microglial migration ability in neuropsychological disorders by regulating the expression of inducible NOS. Results from this study might lead to the identification of novel targets for the treatment of neuropsychological diseases, including neuroinflammatory diseases, stroke, and neurodegenerative diseases.
基金supported by National Natural Science Foundation of China(31801308)Henan Provincial Higher Education Key Research Project(21A210024)CMA·Henan Key Laboratory of Agrometeorological Support and Applied Technique(AMF202109)。
文摘Abiotic stress such as high temperature at flowering is one of many conditions reducing yield of corn(Zea mays L.).Mixing corn cultivars with diverse functional traits increases within-crop diversity and provides a potential means of mitigating yield losses under stress conditions.We conducted a three-year field study to investigate the effects of cultivar mixtures on kernel setting rate,pollen sources,and yield.This study consisted of six treatments,including two high temperature-tolerant(HTT)monocrops of WK702 and DH701,two high temperature-sensitive(HTS)monocrops of DH605 and DH662,and two HTT–HTS mixtures of WK702-DH605 and DH701-DH662.The anthesis–silking interval(ASI)was 0.9–1.6 days shorter in mixtures than in monocrops.Kernel setting rate was increased in mixtures(86.4%–88.7%)compared with those in monocrops(74.7%–84.1%)as a result of synchrony and complementarity of pollination.Grain yields of the HTT–HTS mixtures increased by 13.3%–18.7%,equivalent to 1169 to1605 kg ha^(-1),in comparison with HTS corn monocrops.The results of SSR markers showed that crossfertilization percentage in corn cultivar mixtures ranged from 29.3%to 47.8%,partially explaining yield improvement.Land equivalent ratio(LER)was 1.12 for corn mixtures and the partial land equivalent ratio(e.g.,>0.5)showed the complementary benefits in corn mixtures.The results indicated that mixing corn cultivars with diverse flowering and drought-tolerance traits increased yields via pollination synchrony.
文摘Due to its high efficiency, high precision and high flexibility, CNC lathe is widely used in the machinery manufacturing industry increasingly, and becomes one of CNC machine too[s that most widely used. However, to give full play to the role of CNC lathes, the key is programme, that is, preparing the reasonable and efficient processing procedures depending on the features and precision parts. This paper discussed the problems of programming and processing techniques of the CNC lathe parts.
基金supported by the China Agriculture Research System(CARS-02-20)the Henan Province Agro-ecosystem Field Observation and Research Station,China(30602535)。
文摘Waterlogging stress significantly impairs plant growth and reduces crop yields.Spermidine(Spd),functioning as a second messenger,demonstrates positive effects on plant growth under waterlogging stress conditions.However,the molecular mechanisms by which exogenous Spd application alleviates waterlogging stress remain unclear.This study employed physiological analysis and multi-omics approaches to investigate the effect of Spd application on waterlogging stress.The application of Spd enhanced the expression of genes related to light-harvesting complex(LHC),photosynthesis,and starch-related pathways,while inhibiting chlorophyll degradation and maintaining higher photosynthetic rates,thereby increasing biomass accumulation under waterlogging stress.The activation of genes associated with trehalose and Spd biosynthesis resulted in elevated accumulation of trehalose and endogenous Spd.The inhibition of 1-aminocyclopropane-1-carboxylic acid(ACC)oxidase(ACO)expression contributed to reduced ethylene emission,enhancing maize resistance to waterlogging.Following Spd application,auxin-related genes were up-regulated and indole acetic acid(IAA)content increased,promoting cell elongation in maize and maintaining normal growth under waterlogging stress.Additionally,the upregulation of lipid-related genes led to increased lipid content,protecting cell membranes under waterlogging conditions.These molecular and physiological modifications collectively enhanced resistance to waterlogging stress.These findings advance our understanding of Spd's regulatory roles in mitigating waterlogging damage and provide valuable insights for breeding waterlogging-tolerant maize varieties.
基金funded by the National Natural Science Foundation of China(62273213,62472262,62572287)Natural Science Foundation of Shandong Province(ZR2024MF144)+1 种基金Natural Science Foundation of Shandong Province for Innovation and Development Joint Funds(ZR2022LZH001)Taishan Scholarship Construction Engineering.
文摘Accurately counting dense objects in complex and diverse backgrounds is a significant challenge in computer vision,with applications ranging from crowd counting to various other object counting tasks.To address this,we propose HUANNet(High-Resolution Unified Attention Network),a convolutional neural network designed to capture both local features and rich semantic information through a high-resolution representation learning framework,while optimizing computational distribution across parallel branches.HUANNet introduces three core modules:the High-Resolution Attention Module(HRAM),which enhances feature extraction by optimizing multiresolution feature fusion;the Unified Multi-Scale Attention Module(UMAM),which integrates spatial,channel,and convolutional kernel information through an attention mechanism applied across multiple levels of the network;and the Grid-Assisted Point Matching Module(GPMM),which stabilizes and improves point-to-point matching by leveraging grid-based mechanisms.Extensive experiments show that HUANNet achieves competitive results on the ShanghaiTech Part A/B crowd counting datasets and sets new state-of-the-art performance on dense object counting datasets such as CARPK and XRAY-IECCD,demonstrating the effectiveness and versatility of HUANNet.
基金supported by the National Key R&D Program of China(2019YFA0904000)the National Natural Science Foundation of China(31570094,81502962,32270088)+6 种基金the 111 Project(B16030)the Shan-dong Provincial Natural Science Foundation of China(ZR2020MC015,ZR2018ZC2261)the Taishan Scholar Program of Shandong Provincethe Fundamental Research Funds of Shandong University(2018GN021)the Open Project Program of the State Key Laboratory of Bio-based Material and Green Papermaking(KF201825)the Science and Technology Project of Hunan Province(2021NK1040)Natural Science Foundation of Changsha(KQ2208130).
文摘Bacillus subtilis plays an important role in fundamental and applied research,and it has been widely used as a cell factory for the production of enzymes,antimicrobial materials,and chemicals for agriculture,medicine,and industry.However,genetic manipulation tools for B.subtilis have low efficiency.In this work,our goal was to develop a simple recombineering system for B.subtilis.We showed that genome editing can be achieved in B.subtiliis 1A751 through co-expression of YqaJ/YqaK,a native phage recombinase pair found in B.sub-tilis 168,and the competence master regulator ComK using a double-stranded DNA substrate with short ho-mology arms(100 bp)and a phosphorothioate modification at the 5′-end.Efficient gene knockouts and large DNA insertions were achieved using this new recombineering system in B.subtilis 1A751.As far as we know,this is the first recombineering system using the native phage recombinase pair YqaJ/YqaK in B.subtilis.In conclusion,this new recombineering system provides a simple and fast tool for genetic manipulation of B.sub-tilis,and it will promote studies of genome function,construction of production strains,and genome mining in B.subtilis.
