Expression of Anti-Lipopolysaccharide Factor Isoform 3 in Chlamydomonas reinhardtii Showing High Antimicrobial Activity     

Anguo Li  Ruihao Huang  Chaogang Wang  Qunju Hu  Hui Li  Xiao Li 《Marine drugs》2021,19(5)

Antimicrobial peptides are a class of proteins with antibacterial functions. In this study, the anti-lipopolysaccharide factor isoform 3 gene (ALFPm3), encoding an antimicrobial peptide from Penaeus monodon with a super activity was expressed in Chlamydomonas reinhardtii, which would develop a microalga strain that can be used for the antimicrobial peptide production. To construct the expression cluster, namely pH2A-Pm3, the codon optimized ALFPm3 gene was fused with the ble reporter by 2A peptide and inserted into pH124 vector. The glass-bead method was performed to transform pH2A-Pm3 into C. reinhardtii CC-849. In addition to 8 μg/mL zeocin resistance selection, the C. reinhardtii transformants were further confirmed by genomic PCR and RT-PCR. Western blot analysis showed that the C. reinhardtii-derived ALFPm3 (cALFPm3) was successfully expressed in C. reinhardtii transformants and accounted for 0.35% of the total soluble protein (TSP). Furthermore, the results of antibacterial assay revealed that the cALFPm3 could significantly inhibit the growth of a variety of bacteria, including both Gram-negative bacteria and Gram-positive bacteria at a concentration of 0.77 μM. Especially, the inhibition could last longer than 24 h, which performed better than ampicillin. Hence, this study successfully developed a transgenic C. reinhardtii strain, which can produce the active ALFPm3 driven from P. monodon, providing a potential strategy to use C. reinhardtii as the cell factory to produce antimicrobial peptides.  相似文献   

The role of ear environment in postharvest susceptibility of maize to toxigenic Aspergillus flavus     

Samuel K. Mutiga  Nelson Chepkwony  Owens A. Hoekenga  Sherry A. Flint‐Garcia  Rebecca J. Nelson 《Plant Breeding》2019,138(1):38-50

A kernel screening assay (KSA) was used to assess the genetic and environmental effects on the vulnerability of maize to aflatoxin accumulation. Kernels of 26 inbred lines that had been grown in seven environments, and 190 lines of the Intermated B73xMo17 (IBM) population grown in one location in the United States, were inoculated with a toxigenic strain of A. flavus and incubated in the dark at 30°C for 6 days. Percent kernel colonization (PKC), sporulation and aflatoxin were influenced by the maize genotypes (G), the location (“ear environment” or E) and the GxE interactions. Overall, low broad‐sense heritabilities were observed for PKC, sporulation and aflatoxin. PKC was significantly correlated with sporulation in all environments. Aflatoxin was positively correlated with colonization for two and with sporulation for all ear environments. Higher grain sulphur or magnesium in IBM was associated with less colonization or aflatoxin. Postharvest susceptibility of maize to aflatoxin is thus influenced by factors that are modulated by the ear environment. In a KSA, sporulation could be a proxy test for aflatoxin accumulation.  相似文献   

应用FA及PPA-ELISA技术对猪瘟的检测   总被引：3，自引：0，他引：3  

潘树德  何利昆  李学俭  马兴元 《动物医学进展》2003,24(5):114-116

用免疫荧光抗体诊断技术及单克隆抗体纯化酶联免疫吸附试验诊断技术对疑为猪瘟病毒感染猪进行了检测 ,重点阐述了 2种方法的技术原理和操作过程。通过用免疫荧光抗体诊断技术检测了 5份病料 ,其中有 2份为阳性 ,阳性率为 40 % ;用单克隆抗体纯化酶联免疫吸附试验诊断技术检测了猪瘟血清抗体 ,在 40头母猪血清样品中 ,猪瘟弱毒抗体效价 OD值较高 ,有 1 0 0 %的保护率 ,但是发现母猪群中有 1 0 %隐性猪瘟感染 ,其强毒抗体效价 OD值大于 0 .5,体内带有猪瘟病毒。结果及过程表明此两种诊断方法检测快速、鉴别准确、分辨率高  相似文献   

马传贫驴白细胞弱毒疫苗株跨膜蛋白主要免疫决定区基因的克隆与表达   总被引：3，自引：0，他引：3  

薛飞  贾斌  朱远茂  杨建德  王晓钧  相文华  赵立平  吕晓玲  沈荣显 《中国预防兽医学报》2003,25(5):321-325

从感染驴白细胞的马传贫驴白细胞弱毒疫苗株前病毒DNA中克隆了编码跨膜蛋白主要免疫决定区(TMIR)的基因，并在大肠杆菌中进行了表达。所表达的融合蛋白有一部分是可溶的，其氨基端带有6个组氨酸的标签，因此可以用固定化金属离子亲和层析法在非变性条件下进行纯化。在间接酶联免疫吸附试验(ELISA)和免疫印迹试验中，重组的TMIR蛋白可与马传贫阳性血清样品发生反应，而与健康马血清无任何反应。这表明该重组蛋白具有良好的抗原性和特异性，可用于马传贫弱毒疫苗株在体内外复制、接种马体内免疫应答及马传贫诊断的研究。  相似文献   

表面技术在内燃机气缸套中的新进展     

李梅广孙宏飞  赵剑波孔红杰 《农业装备与车辆工程》2005,(3):41-44

论述了气缸套的失效机理，针对这一缺点，较全面地阐述了当前表面技术在气缸套上的应用现状，并提出了未来气缸套的发展方向。为同行提供了参考和启迪，也为拓宽热喷涂技术的应用指明了一个方向。  相似文献   

Surgical repair of skull fractures in four horses using cuttable bone plates.     

B A Dowling  A J Dart  G Trope 《Australian veterinary journal》2001,79(5):324-327

Three horses with severely comminuted, open facial bone fractures and one horse with a comminuted, open orbital rim fracture were referred for treatment. Severe facial bone asymmetry and epistaxis were apparent in all cases and subcutaneous emphysema was present in two, however physical and neurological examinations were otherwise normal. Radiography and endoscopy were of some use in assessing the degree of damage, although the true extent of the damage was more apparent at surgery. Surgical reduction of the fractures was recommended to maximise cosmetic and functional outcome. After surgical debridement and reduction of the fractures the bone fragments remained unstable and were not amenable to stabilisation with interfragmentary wires alone, so 2 mm cuttable bone plates were used to maintain fracture alignment. Screw migration occurred in three horses and in one of these horses the plate had to be removed. Other complications were minor and in all horses the fractures healed with good cosmetic and functional outcome. The use of cuttable bone plates should be considered as a reasonable alternative to inter-fragmentary wiring for unstable, comminuted fractures of the facial bones, even where fractures are open.  相似文献   

木瓜剖分原理研究及新机型设计     

王晓光  彭斌  陈宏  陈明森 《农机化研究》2009,31(1)

针对我国目前对木瓜剖半设备研究较少,剖切方式不合理,切割效果差等问题,设计开发了新型木瓜剖半机。该机切割方式选用滑切切割的方式,由运料系统、切割系统、出料系统组成,设计合理,结构简单。为此,计算了滑切角、切割角等主要切割参数,并给出了新型剖半机的主要技术参数。试验表明:该机切割效果好,能够有效提高生产效率。  相似文献   

Neutrophil adherence,phagocytic-nitroblue tetrazolium reduction and chemiluminescence in canine whole blood     

H. Nagahata  T. Sako  J. A. Reiter  S. P. Dibartola  C. G. Couto  G. J. Kociba 《Veterinary research communications》1991,15(3):181-188

Methods for measuring neutrophil adherence, phagocytic-nitroblue tetrazolium (NBT) reducing activity and chemiluminescence were applied to canine whole blood as means for routine assessment of neutrophil functions. The phagocytic-NBT reduction test appeared to be useful for monitoring the NBT reducing activity of phagocytic cells associated with phagocytic functions. Ethylene diamine tetraacetic acid suppressed both the adherence and the phagocytic-NBT reducing activity of neutrophils. Increased phagocytic-NBT reduction and an enhanced chemiluminescence response were observed in dogs with neutrophilia. These methods provide a rapid and practical screening procedure for measuring selected phagocytic functions in canine whole blood.  相似文献   

用酶联免疫吸附试验检测鸡白血病以及控制、根除鸡白血病措施的探讨     

李厚达 《扬州大学学报(农业与生命科学版)》1987,(4)

用酶联免疫吸附试验(ELISA),对2群鸡的鸡蛋清和1株鸡的马立克氏病疫苗进行检测,发现2群鸡的鸡蛋清中,鸡白血病病毒的阳性率分别是11％和29％,鸡马立克氏病冻干苗隐藏鸡白血病病毒群体特异性(gs)抗原的阳性率为100％。本文指出我国禽苗可能带有鸡的白血病病毒,分析讨论了鸡白血病病毒的垂直传递和水平传播的规律,提出在曾祖代和祖代鸡群中,采用ELISA试验,检测鸡蛋清,能减少以至根除鸡的白血病。  相似文献   

Development of a semi-selective medium and an immunofluorescence colony-staining procedure for the detection of Clavibacter michiganensis subsp. sepedonicus in cattle manure slurry     

N. J. M. Roozen  J. W. L. Van Vuurde 《European journal of plant pathology / European Foundation for Plant Pathology》1991,97(5):321-334

Various compounds and basal media were tested for their suitability to create a semi-selective medium for isolation ofClavibacter michiganensis subsp.sepedonicus (Cms) from cattle manure slurry containing c. 10⁸ colony forming units (cfu) per ml.Plating efficiency of Cms in yeast glucose mineral medium (YGM) was 104% compared with yeast peptone glucose medium. Nalidixic acid, polymyxin B sulphate and the experimental disinfectant S-0208 inhibited colony growth of cattle slurry bacteria as compared with Cms in YGM. The optimal concentration of these inhibitors in combination was determined by modified agar diffusion tests and by pour plating in 24-well tissue culture plates. The semi-selective medium YGMI consisted of YGM supplemented with nalidixic acid (2 mg/l), polymyxin B sulphate (30 mg/l) and S-0208 (125 mg/l). Plating efficiency varied for Cms between 50.9 and 69.6%, for cattle slurry bacteria between 1.8 and 2.5% and for saprophytes from potato heel end extracts between 11.5 and 27.4%.Differentiation of Cms colonies from other colonies was based on their small and bluish colony morphology in pour plates and on immunofluorescence colony-staining (IFC). IFC of a pure culture of micro colonies of Cms in YGM was possible after one day incubation (colonies c. 5 cells). Green background fluorescence in the agar gels was prevented by addition of Tween 20 (0.1%) to the washing buffer and the use of 1% agar gels. IFC of macro colonies of Cms in YGMI, visible with 4x objective magnification, was possible after 4 days. The detection level of the target organism in artificially inoculated cattle slurry in YGMI based on colony morphology varied between 1.4×10³ and 2.3×10⁴ cfu per ml of cattle slurry. Miniaturized plating combined with IFC, using wells in tissue culture plates (=16 mm), proved suitable for detection, but was c. 30 times les sensitive. The recovery of Cms was negatively correlated with the number of saprophytic colonies in the agar plates (R ²=0.74).  相似文献