The NAC(NAM,ATAF,and CUC)superfamily is one of the largest plant-specific families containing transcription factors.An increasing number of studies suggest that NAC1 is involved in plants response to different biotic ...The NAC(NAM,ATAF,and CUC)superfamily is one of the largest plant-specific families containing transcription factors.An increasing number of studies suggest that NAC1 is involved in plants response to different biotic and abiotic stimulis.Nicotiana benthamiana is a widely used system for evaluating plant-pathogen interactions.In order to study the biochemical function of NbNAC1,NbNAC1 protein and antibody are essential.Therefore,we focused on developing a prokaryotic expression system for producing the Nicotiana benthamiana NbNAC1 protein of in Escherichia coli and the preparation of its polyclonal antibody.Firstly,we constructed two different molecular weight prokaryotic expression vectors:pGE vector with GST tag(pGEX4T-1–NbNAC1)and pET expression vector with His tag(pET28a-NbNAC1).The NbNAC1 protein can be successfully expressed in both vectors.The His-tagged NbNAC1 proteins are insoluble,while the GST-tagged NbNAC1 proteins are partially soluble.We then successfully purified and enriched both proteins.The His-tagged NbNAC1 was chosen to immunize rabbits owing to an unknown protein accompanying the GST-tagged NbNAC1.The anti-NbNAC1 polyclonal antibody had good specificity and could be used in subsequent protein-related studies.展开更多
Sonodynamic therapy(SDT)has attracted widespread interest in biomedicine,owing to its novel and noninvasive therapeutic method triggered by ultrasound(US).Herein,the Ti_(3)C_(2) MXene nanosheets(Ti_(3)C_(2) NSs)are de...Sonodynamic therapy(SDT)has attracted widespread interest in biomedicine,owing to its novel and noninvasive therapeutic method triggered by ultrasound(US).Herein,the Ti_(3)C_(2) MXene nanosheets(Ti_(3)C_(2) NSs)are developed as good sonosensitizers via a two-step method of chemical exfoliation and high-temperature treatment.With the high-temperature treatment,the oxygen defect of Ti_(3)C_(2) MXene nanosheets(H-Ti_(3)C_(2) NSs)is greatly increased.Therefore,the electron(e^(-))and hole(h^(+))generated by US can be separated faster due to the improved degree of oxidation,and then the recombination of e^(-)-h^(+)can be prevented with the abundant oxygen defect under US irradiation,which induced the sonodynamic efficiency greatly to improve around 3.7-fold compared with Ti_(3)C_(2) NSs without high-temperature treatment.After PEGylation,the H-Ti_(3)C_(2)-PEG NSs show good stability and biocompatibility.In vitro studies exhibit that the inherent property of mild photothermal effect can promote the endocytosis of H-Ti_(3)C_(2)-PEG NSs,which can improve the SDT efficacy.In vivo studies further display that the increased blood supply by the mild photothermal effect can significantly relieve hypoxia in the tumor microenvironment,showing photothermal therapy(PTT)enhanced SDT.Most importantly,the H-Ti_(3)C_(2)-PEG NSs can be biodegraded and excreted out of the body,showing no significant long-term toxicity.Our work develops the defective H-Ti_(3)C_(2) NSs as high-efficiency and safe sonosensitizers for photothermal-enhanced SDT of cancer,extending the biomedical application of MXene-based nanoplatforms.展开更多
基金This work was supported by the National Natural Science Foundation of China(31500209)Natural Science Foundation of Jiangsu Province of China,(BK20201431)+2 种基金Agricultural science and technology independent innovation Foundation of Jiangsu Province of China(CX(20)3128)Open Project Program of Joint International Research Laboratory of Agriculture and Agri-Product Safety,the Ministry of Education of China,Yangzhou University(JILARKF202006)Qing Lan Project of Yangzhou University and Yangzhou University of“High-end Talent Support Program”.
文摘The NAC(NAM,ATAF,and CUC)superfamily is one of the largest plant-specific families containing transcription factors.An increasing number of studies suggest that NAC1 is involved in plants response to different biotic and abiotic stimulis.Nicotiana benthamiana is a widely used system for evaluating plant-pathogen interactions.In order to study the biochemical function of NbNAC1,NbNAC1 protein and antibody are essential.Therefore,we focused on developing a prokaryotic expression system for producing the Nicotiana benthamiana NbNAC1 protein of in Escherichia coli and the preparation of its polyclonal antibody.Firstly,we constructed two different molecular weight prokaryotic expression vectors:pGE vector with GST tag(pGEX4T-1–NbNAC1)and pET expression vector with His tag(pET28a-NbNAC1).The NbNAC1 protein can be successfully expressed in both vectors.The His-tagged NbNAC1 proteins are insoluble,while the GST-tagged NbNAC1 proteins are partially soluble.We then successfully purified and enriched both proteins.The His-tagged NbNAC1 was chosen to immunize rabbits owing to an unknown protein accompanying the GST-tagged NbNAC1.The anti-NbNAC1 polyclonal antibody had good specificity and could be used in subsequent protein-related studies.
基金partially supported by the National Key Research and Development Program of China(2022YFB3804600 and 2021YFF0701800)the National Natural Science Foundation of China(U20A20254 and 52072253)+7 种基金Collaborative Innovation Center of Suzhou Nano Science and TechnologySuzhou Key Laboratory of Nanotechnology and Biomedicinethe Jiangsu Natural Science Fund for Distinguished Young Scholars(BK20211544)the 111 ProjectJoint International Research Laboratory of Carbon-Based Functional Materials and DevicesMedical Research Project of Jiangsu Province(ZD2022024)New Cornerstone Science Foundation through the XPLORER PRIZEKey Laboratory of Structural Deformities in Children of Suzhou(SZS2022018)。
基金partially supported by the National Research Programs of China(2016YFA0201200)the National Natural Science Foundation of China(U20A20254,52072253)+3 种基金Collaborative Innovation Center of Suzhou Nano Science and Technology,a Jiangsu Social Development Project(BE2019658)a Project Funded by the Priority Academic Program Development(PAPD)of Jiangsu Higher Education Institutionssupported by the Tang Scholarship of Soochow Universitythe fundamental Research Funds for Central Universities(2662019PY024).
文摘Sonodynamic therapy(SDT)has attracted widespread interest in biomedicine,owing to its novel and noninvasive therapeutic method triggered by ultrasound(US).Herein,the Ti_(3)C_(2) MXene nanosheets(Ti_(3)C_(2) NSs)are developed as good sonosensitizers via a two-step method of chemical exfoliation and high-temperature treatment.With the high-temperature treatment,the oxygen defect of Ti_(3)C_(2) MXene nanosheets(H-Ti_(3)C_(2) NSs)is greatly increased.Therefore,the electron(e^(-))and hole(h^(+))generated by US can be separated faster due to the improved degree of oxidation,and then the recombination of e^(-)-h^(+)can be prevented with the abundant oxygen defect under US irradiation,which induced the sonodynamic efficiency greatly to improve around 3.7-fold compared with Ti_(3)C_(2) NSs without high-temperature treatment.After PEGylation,the H-Ti_(3)C_(2)-PEG NSs show good stability and biocompatibility.In vitro studies exhibit that the inherent property of mild photothermal effect can promote the endocytosis of H-Ti_(3)C_(2)-PEG NSs,which can improve the SDT efficacy.In vivo studies further display that the increased blood supply by the mild photothermal effect can significantly relieve hypoxia in the tumor microenvironment,showing photothermal therapy(PTT)enhanced SDT.Most importantly,the H-Ti_(3)C_(2)-PEG NSs can be biodegraded and excreted out of the body,showing no significant long-term toxicity.Our work develops the defective H-Ti_(3)C_(2) NSs as high-efficiency and safe sonosensitizers for photothermal-enhanced SDT of cancer,extending the biomedical application of MXene-based nanoplatforms.
