气肿疽梭菌CctA基因的原核表达及其间接ELISA检测方法的建立     

李新苹  陈晓洁  赵兵  刘强德  丁占强  张飞  唐慧芬  曹剑  侯凤  刘洋  王钢 《中国畜牧兽医》2021,48(5):1876-1882

研究旨在原核表达气肿疽梭菌细胞毒素A （CctA）基因,用纯化的重组蛋白建立其间接ELISA检测方法,a。利用大肠杆菌密码子的偏爱性优化CctA基因序列,克隆至原核表达载体pET-28a （+）,双酶切鉴定原核表达质粒pET28a-CctA并测序。将重组质粒转入大肠杆菌,经IPTG诱导得到高表达的重组CctA包涵体蛋白。包涵体蛋白变性后经镍（Ni）柱纯化,SDS-PAGE检测其纯化效果。以豚鼠抗气肿疽梭菌抗血清为一抗,用Western blotting方法检测重组CctA蛋白的反应原性。用棋盘滴定法建立间接ELISA检测方法,以复性后的重组CctA蛋白作为检测抗原,摸索抗原包被浓度、封闭液的种类及浓度、抗体的最适稀释度和反应条件等。选用3批气肿疽灭活疫苗免疫豚鼠后采集血清,同时用攻毒和间接ELISA两种方法验证疫苗的免疫效力。质粒双酶切结果显示,得到大小约853 bp的条带,与预期相符,且测序结果正确。SDS-PAGE结果表明,成功表达并纯化大小为35 ku的重组CctA蛋白。Western blotting结果显示,豚鼠抗气肿疽梭菌抗血清与重组CctA蛋白具有良好的反应原性。建立的间接ELISA法的最适条件为：抗原的包被浓度为0.5 μg/mL,于4 ℃包被过夜;封闭液选择10%胎牛血清,37 ℃孵育2 h;二抗的稀释度为1：8 000;室温避光显色10 min后终止反应,测定D_{450 nm}值。当P/N>4.6时,间接ELISA法检测结果与豚鼠攻毒试验结果拟合度较好。本研究成功表达CctA基因并纯化了重组CctA蛋白,建立的以重组CctA蛋白为检测抗原的间接ELISA检测方法,有望成为气肿疽灭活疫苗免疫效果验证的替代方法。  相似文献   

基于HYDRUS-2D的地下滴灌下水分运移数值模拟研究   总被引：2，自引：0，他引：2  

潘红霞  付恒阳  贺屹 《灌溉排水学报》2015,34(3)

利用HYDRUS-2D软件模拟了地下滴灌条件下滴头周围黏壤土的土壤水分动态,并与田间观测进行对比;通过分析不同滴头流量对试验地土壤湿润模式的影响,明确了最佳灌水技术参数。结果表明,HYDRUS-2D软件模拟结果与观测值一致,体积含水率均方根误差为1.2%~4.5%,湿润范围均方根误差变化范围为2.1~3.87cm。  相似文献   

生物杀虫剂组合在有机水稻种植中的作用效果     

刘蓉#  赵薇#  农向群  周燕  石育萍  于丽  杨金娟  王广君  涂雄兵  张泽华 《植物保护》2023,49(3):310-316

针对上海市崇明区有机水稻生产过程中病虫害防治效果不佳的问题,在水稻无化肥、无化学农药种植条件下开展了生物杀虫剂对水稻主要害虫稻纵卷叶螟和稻飞虱的田间防控效果评价。在崇明区的2个试验基地,于水稻不同生长时期,以绿僵菌、白僵菌产品分别与病毒杀虫剂MbNPV、植物源杀虫剂苦参碱和细菌杀虫剂Bt组合进行施药处理。结果表明,应用绿僵菌/白僵菌与MbNPV组合,能够有效防治稻纵卷叶螟,在稻纵卷叶螟发生较早的基地,施药后7 d和14 d的防效可达58.2%～81.2%,而在稻纵卷叶螟发生较晚的另一基地,施药后7 d防效可达61.3%和65.6%;绿僵菌/白僵菌与苦参碱组合,可用于防治稻飞虱,7 d和14 d的防效达64.7%～76.1%;绿僵菌/白僵菌与苏云金杆菌组合防治稻纵卷叶螟的14 d防效分别达64.0%和75.4%,优于对稻飞虱的防效;各组合试验结果显示,白僵菌有相对较长的持效作用,且在害虫迁入早期阶段(水稻发育分蘖盛期至拔节期)施用生物农药比在中后期(抽穗期及之后)施用的防效高。可见,生物杀虫剂组合可作为农作物绿色防控的有效手段。  相似文献   

Acute oral toxicity of zinc phosphide: an assessment for wild house mice (Mus musculus)     

Lyn A. HINDS  Steve HENRY  Nikki VAN DE WEYER  Freya ROBINSON  Wendy A. RUSCOE  Peter R. BROWN 《Integrative zoology》2023,18(1):63-75

Irregular plagues of house mice, Mus musculus, incur major economic impacts on agricultural production in Australia. The efficacy of zinc phosphide (ZnP), the only registered broadacre control agent for mice, is reported as increasingly variable. Have mice become less sensitive over time or are they taking a sub-lethal dose and developing aversion? In this laboratory study, the sensitivity of mice (wild caught; outbred laboratory strain) was assessed using oral gavage of a range of ZnP concentrations. The estimated LD₅₀ values (72–79 mg ZnP/kg body weight) were similar for each mouse group but are significantly higher than previously reported. The willingness of mice to consume ZnP-coated grains was determined. ZnP-coated grains (50 g ZnP/kg grain) presented in the absence of alternative food were consumed and 94% of wild mice died. Mice provided with alternative food and ZnP-coated wheat grains (either 25 or 50 g ZnP/kg grain) consumed toxic and non-toxic grains, and mortality was lower (33–55%). If a sublethal amount of ZnP-coated grain was consumed, aversion occurred, mostly when alternative food was present. The sensitivity of wild house mice to ZnP in Australia is significantly lower than previously assumed. Under laboratory conditions, ZnP-coated grains coated with a new higher dose (50 g ZnP/kg grain) were readily consumed. Consumption of toxic grain occurred when alternative food was available but was decreased. Our unambiguous findings for house mice indicate a re-assessment of the ZnP loading for baits used for control of many rodents around the world may be warranted.  相似文献   

干旱区小麦多功能高分子植物生长调节剂增产和品质改善试验研究   总被引：3，自引：0，他引：3  

张松林  赵首彩  董庆士  田侠  房妮 《中国农学通报》2005,21(9):263-265

以观测记录和拍照方式研究多功能高分子植物生长调节剂对小麦生长和繁殖、品质的影响,分别用土壤含水量为试剂保水指标,小麦生长、结实和产量指标、小麦品质指标以及经济效益指标与对照比较以评定试验效果。结果显示,施用试剂能够提高土壤保水率,有利于小麦的营养和结实生长,使小麦产量提高12.30%。小麦粗蛋白、赖氨酸和湿面筋含量均有显著提升,不利于人体健康的灰分和粗脂肪含量显著降低;经济效益有所提高,仅仅考虑增产就净增产值734.25￣794.85元/hm2。  相似文献   

VISAR Data Reducing Method and Its Application to Impact Experiment     

YAO Guo-wen  LIU Zhan-fang 《保鲜与加工》2006,(8):25-28

Identification and adjustment on the disfigurements of original data of VISAR are probed by developing software for VISAR data reducing based on measure theory. Then the software is applied to plate impact experiment of ceramics. The disfigurements of light intensity data are identified and adjusted through Lissajou~ figures. The results indicate that VISAR data are reduced quickly and effectively by the software, and the exact free surface velocity history is obtained.  相似文献   

Experiment for Condensing Heat Transfer Performance in a New Type of Horizontal Three Dimensional Inner Microfin Tubes     

Du Yang Xin Mingdao Huang Shaoming 《保鲜与加工》1995,(5):50-56

A new type of three dimenstional inner microfin tube is devoloped. Under four different pressures and in the range of condensing mass velocities from 33 to 153 kg/m2 . s,and by the experimental study of the three kinds of tubes with different microfin parameters, the performances of the condensation heat transfer for R11 in the horizontal microfin tubes and obtained. The experiments show that the inner microfin tube is a hopeful and nighty effective condensing heat transfer enhancement tube.  相似文献   

Study of Combined Heat Transfer Enhancement with Swirl Flow Generator and Flag-Type Insents     

Chen Li  Xiang Dong 《保鲜与加工》1996,(1):64-67

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The Imitate Experiment and Research of Raft Floating Breakwater     

《保鲜与加工》2003,(1):44-48

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Study on Mechanism of the Reinforcing Effect of Geogrid in Soil Dam     

Wen ShuLian  Wang KeZhong 《保鲜与加工》2005,(3):68-71

An experimental study on consolidation and drainage of two gravels with different grading was carried out using high - pressure large - scale tri - axial testing facility. A concept of equivalent surrounding pressure is introduced. An analysis was carried out on the effect and mechanism of reinforcement, experimental surrounding pressure and soil properties. Results show that the reinforcement can improve the strength and failure strain of the material and delay the failure of the soil. The effect of reinforcement of soil - gravel depends on the surrounding pressure and weathering of the gravel.  相似文献