丝裂原活化蛋白激酶15(mitogen-activated protein kinase 15,MAPK15),又称ERK7或ERK8,是MAPK家族的非典型新成员。MAPK15不同程度地促进不同肿瘤细胞的增殖、迁移、自噬等细胞活动。本研究以MAPK15为靶点,筛选特异性的MAPK15纳米抗体,...丝裂原活化蛋白激酶15(mitogen-activated protein kinase 15,MAPK15),又称ERK7或ERK8,是MAPK家族的非典型新成员。MAPK15不同程度地促进不同肿瘤细胞的增殖、迁移、自噬等细胞活动。本研究以MAPK15为靶点,筛选特异性的MAPK15纳米抗体,评估其是否能够作为免疫组织化学和Western印迹中的一抗用于其抗原表达的检测,并探究该纳米抗体在B16-F10黑色素瘤细胞中的作用。通过噬菌体展示技术从B16-F10黑色素瘤细胞纳米抗体文库中进行筛选,得到1株MAPK15特异性纳米抗体,命名为MAPK15-VHH;将该菌株构建原核表达载体,进行优化诱导表达条件时发现,0.6 nmol/L IPTG,15℃,100 r/min条件下该纳米抗体的上清表达量最高。通过竞争ELISA法检测MAPK15-VHH的亲和力,结果显示,该抗体KD值为0.9829。通过Western印迹和免疫组织化学检测脑组织中MAPK15在蛋白质水平的表达量及分布情况,结果表明,MAPK15-VHH可与组织中的MAPK15结合,用于检测MAPK15蛋白的表达情况。在B16-F10黑色素瘤细胞中过表达MAPK15后向其中添加MAPK15-VHH,通过CCK8、Western印迹以及实时荧光定量PCR检测其对该细胞增殖和自噬的影响,结果显示,该MAPK15-VHH能作为MAPK15的拮抗剂,抑制B16-F10细胞的增殖和自噬,从而抑制黑素瘤细胞的生长。综上所述,本研究成功得到一株亲和力较强的MAPK15纳米抗体,可用于Western印迹及免疫组织化学以检测MAPK15在蛋白质水平的表达及分布;此外,该MAPK15纳米抗体可以作为MAPK15拮抗剂,有效地抑制B16-F10细胞的增殖和自噬进程,为临床检测试剂和治疗药物的开发提供理论基础。展开更多
The literature reports mainly focus on 2023-2024,and the research hotspots are mostly productivity and new quality productivity.After analysis,research in the field of fisheries is still in its early stages.Among them...The literature reports mainly focus on 2023-2024,and the research hotspots are mostly productivity and new quality productivity.After analysis,research in the field of fisheries is still in its early stages.Among them,the facilities,intelligence,and"Blue granary"of marine fisheries need to be further strengthened to provide technical support for achieving the goal of"new quality productivity"in the fishing industry in the future.展开更多
According to the"National Modern Facility Agriculture Construction Plan(2023-2030)released by the Ministry of Agriculture and Rural Affairs,the"Special Implementation Plan for Modern Facility Fisheries Const...According to the"National Modern Facility Agriculture Construction Plan(2023-2030)released by the Ministry of Agriculture and Rural Affairs,the"Special Implementation Plan for Modern Facility Fisheries Construction"emphasizes the development of circular aquaculture,active expansion of green aquaculture space in facility fisheries,multi-channel development of food sources,comprehensive improvement of the quality,efficiency,and competitiveness of facility fisheries,ensuring stable and safe supply of aquatic products,and better meeting people's needs for high-quality aquatic products and beautiful aquatic ecological environment.Since the implementation of the 14th Five Year Plan for fisheries,facility fisheries have made significant progress.In order to thoroughly implement General Secretary Xi Jinping's important discourse on new quality productivity,conduct in-depth research on the current situation and existing problems of the fishery industry,and put forward suggestions to meet the requirements of high-quality development of the fishery industry in the new era.Therefore,it is crucial to continue practicing the"revitalization of aquatic seed industry"and"big food concept",take multiple measures to"demand food from rivers,lakes,and seas",and expand aquaculture space.Through research,it has been found that research in the field of facility based fisheries is still in its developmental stage.Further exploration is needed on the current status,breeding models,and experimental demonstrations of land-based facility based fisheries,in order to provide a basis for fisheries regulatory authorities to formulate the"15th Five Year Plan"for facility based fisheries.展开更多
基金supported by the Technical system of national bulk freshwater fish industry(No.CARS-45-33)National bulk freshwater fish industry technology system Tianjin freshwater aquaculture industry technology system innovation team(No.ITTFRS2024000)+2 种基金Tianjin Science and Technology Plan Project(No.23YDTPJC00420)Central Guiding Local Science and Technology Development Funds(No.24ZYCGSN00250)Opening Foundation of Ministry of Education Key Laboratory of Pollution Processes and Environmental Criteria(No.202007 and 202105).
文摘The literature reports mainly focus on 2023-2024,and the research hotspots are mostly productivity and new quality productivity.After analysis,research in the field of fisheries is still in its early stages.Among them,the facilities,intelligence,and"Blue granary"of marine fisheries need to be further strengthened to provide technical support for achieving the goal of"new quality productivity"in the fishing industry in the future.
基金supported by the National Bulk Freshwater Fish Industry Technology System(CARS-45-33)Tianjin Freshwater Aquaculture Industry Technology System Innovation Team(ITTFRS20211000-005/002)+2 种基金Tianjin Science and Technology Plan Project(23YDTPJC00420)Central Guiding Local Science and Technology Development Fund(24ZYCGSN00250)Open Fund of Key Laboratory of Environmental Standards and Environmental Pollution of the Ministry of Education(202007,202105).
文摘According to the"National Modern Facility Agriculture Construction Plan(2023-2030)released by the Ministry of Agriculture and Rural Affairs,the"Special Implementation Plan for Modern Facility Fisheries Construction"emphasizes the development of circular aquaculture,active expansion of green aquaculture space in facility fisheries,multi-channel development of food sources,comprehensive improvement of the quality,efficiency,and competitiveness of facility fisheries,ensuring stable and safe supply of aquatic products,and better meeting people's needs for high-quality aquatic products and beautiful aquatic ecological environment.Since the implementation of the 14th Five Year Plan for fisheries,facility fisheries have made significant progress.In order to thoroughly implement General Secretary Xi Jinping's important discourse on new quality productivity,conduct in-depth research on the current situation and existing problems of the fishery industry,and put forward suggestions to meet the requirements of high-quality development of the fishery industry in the new era.Therefore,it is crucial to continue practicing the"revitalization of aquatic seed industry"and"big food concept",take multiple measures to"demand food from rivers,lakes,and seas",and expand aquaculture space.Through research,it has been found that research in the field of facility based fisheries is still in its developmental stage.Further exploration is needed on the current status,breeding models,and experimental demonstrations of land-based facility based fisheries,in order to provide a basis for fisheries regulatory authorities to formulate the"15th Five Year Plan"for facility based fisheries.
