共查询到20条相似文献，搜索用时 46 毫秒
1.
以抗堆型艾美耳球虫子孢子的单抗EASP-3G3作为工具,对鸡各段消化道上皮细胞切片和子孢子进行免疫组化染色,并利用蛋白质印迹技术来检测单抗所识别子孢子可溶性抗原的分子量,来确定子孢子和十二指肠上皮细胞之间是否存在共同抗原。结果表明单抗只与十二指肠上皮细胞发生反应,而与其他肠段无染色反应。而且单抗所识别的可溶性抗原分子量为35~48 ku。抗子孢子的单抗同时与十二指肠上皮细胞和子孢子反应,而不与其他肠段反应,证明堆型艾美耳球虫寄生的位点特异性与十二指肠上皮细胞表面的某种抗原分子有内在的关系。 相似文献
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为了研究鸡柔嫩艾美耳球虫SO7基因原核表达重组蛋白的免疫效力,分析其在鸡抗柔嫩艾美耳球虫感染中对鸡平均增重、抗球虫指数(ACI)、减少盲肠病变和卵囊数量中的作用,从柔嫩艾美耳球虫孢子化卵囊中提取总RNA作为模板,用RT-PCR扩增出SO7基因片段,再应用DNA重组技术将SO7基因克隆到原核表达载体pET32a(+)中并用IPTG诱导表达。SDS-PAGE结果显示,pET32a(+)-SO7表达的目的蛋白约为40ku,主要以包涵体形式存在。用纯化的重组蛋白进行动物免疫试验显示,SO7重组蛋白在鸡抗柔嫩艾美耳球虫感染中有较好的免疫保护作用,能够显著提高鸡柔嫩艾美耳球虫感染鸡的平均增重和抗球虫指数(ACI),对减少盲肠病变和卵囊数量也有部分作用。 相似文献
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抗原虫药主要包括抗球虫药、抗锥虫药、抗梨形虫药、抗滴虫药四大类。其中球虫主要寄生于肠道,其中柔嫩艾美耳球虫主要寄生于盲肠,常见且致病力最强;毒害艾美耳球虫寄生于鸡小肠中段;堆型艾美耳球虫寄生于鸡小肠前段;巨型艾美耳球虫寄生于鸡小肠中段;另外其他种属动物都可 相似文献
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抗原虫药主要包括抗球虫药、抗锥虫药、抗梨形虫药、抗滴虫药四大类.其中球虫主要寄生于肠道,其中柔嫩艾美耳球虫主要寄生于盲肠,常见且致病力最强;毒害艾美耳球虫寄生于鸡小肠中段;堆型艾美耳球虫寄生于鸡小肠前段;巨型艾美耳球虫寄生于鸡小肠中段;另外其他种属动物都可感染球虫病,该病对养鸡和养兔专业户造成经济损失尤其较大. 相似文献
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将已扩增出的鸡堆型艾美耳球虫特异性单抗轻链可变区基因进行纯化,并用纯化的基因片段与pMD-18T载体连接,将重组载体转化于感受态细胞JM109,筛选出阳性重组子,提取阳性重组子质粒并进行测序,得到特异性单抗轻链可变区的基因序列。为鸡堆型艾美耳球虫特异性单链抗体基因的构建奠定了基础。 相似文献
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《中国兽医学报》2017,(5):844-849
本研究将巨型艾美耳球虫Emgam56基因去除信号肽后的ORF序列连接至pET-28a(+)载体,构建表达载体pET-28-Emgam56,转化表达宿主菌大肠杆菌BL21(DE3),经IPTG诱导表达;分析表达产物的可溶性和重组蛋白的分子相对质量,并进一步鉴定重组配子体蛋白rEmGAM56的抗原性。SDS-PAGE分析表明,表达载体能表达56000左右蛋白质,主要以可溶性蛋白的形式存在;Western blot检测表明,rEmGAM56蛋白能被鼠抗rEmGam56多克隆抗体和巨型艾美耳球虫感染鸡的康复血清特异性识别,显示重组蛋白具有良好的抗原性。本研究结果为研制重组配子体抗原亚单位疫苗奠定了基础。 相似文献
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间接血凝检测鸡柔嫩艾美耳球虫血清抗体方法的建立 总被引：1，自引：0，他引：1
《中国兽医杂志》2016,(2)
为了快速检测免疫球虫后宿主血清中特异性抗体的水平,以DE-52层析纯化子孢子、裂殖子,超声裂解获取可溶性蛋白;原核表达EtMIC4-N蛋白,致敏戊二醛醛化后的红细胞,以人工免疫柔嫩艾美耳球虫后分离的血清为阳性血清,以SPF鸡分离的血清为阴性血清,建立柔嫩艾美耳球虫间接血凝试验。结果显示,稀释度为1:2万鞣酸处理戊二醛醛化后的红细胞,使用100μg/mL可溶性裂殖子、子孢子蛋白或12.5μg/mL表达的重组蛋白EtMIC4-N均可以与阳性血清发生反应(可达1:1024),与鸡的阴性血清及大肠杆菌病的阳性血清无反应.与堆型艾美耳球虫、巨型艾美耳球虫阳性血清有微弱反应(不超过1:8)。试验证明,该方法具有简单、方便、快捷、灵敏等优点。 相似文献
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《畜牧与兽医》2020,(11)
提取柔嫩艾美耳球虫(扬州株)配子体基因组DNA,PCR扩增配子体基因Etgam56,经克隆和序列分析后,优化密码子,连接至pET-28a(+),构建原核表达载体pET-28a(+)-Etgam56,优化条件进行体外诱导表达,Western blot分析其抗原性。结果显示,Etgam56基因全长1 425 bp,为一个完整开放阅读框,共编码474个氨基酸;体外表达重组蛋白的大小约为56 ku,IPTG终浓度为0.10 mmol/L,37℃诱导3 h时的表达量最高,且主要以包涵体形式存在。Western blot结果显示,该重组蛋白能被6×HIS标签单克隆抗体、柔嫩艾美耳球虫鸡康复血清和毒害艾美耳球虫鸡康复血清识别,表明该重组蛋白为特异性表达的蛋白,具有很好的抗原性和交叉反应性。本研究将为进一步探析柔嫩艾美耳球虫配子体蛋白的功能与研制鸡球虫基因工程疫苗奠定基础。 相似文献
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旨在探析毒害艾美耳球虫氧化还原酶EnOXIO1的功能。提取毒害艾美耳球虫扬州株配子体总RNA,应用RT-PCR获取EnOXIO1编码基因,构建原核表达载体pET-28a(+)-EnOXIO1,转化至大肠杆菌BL21(DE3),体外诱导表达,对重组蛋白进行抗原性分析,应用制备的多抗进行激光共聚焦免疫荧光定位。结果显示,获得的EnOXIO1基因全长为2 535 bp,体外重组表达的蛋白大小约100 ku,主要以包涵体的形式存在,表达量约为0.5 mg·mL-1。Western blot分析结果显示,该重组蛋白能被6×HIS标签单克隆抗体识别,同时能被制备的鼠多抗以及毒害艾美耳球虫、堆型艾美耳球虫和柔嫩艾美耳球虫病鸡康复血清所识别,表明该重组蛋白具有较好的反应原性和交叉反应原性。激光共聚焦免疫荧光定位结果显示,EnOXIO1基因表达产物主要存在于配子体内的成壁体上,参与了卵囊壁的形成。本研究成功克隆和表达了毒害艾美耳球虫EnOXIO1蛋白,并将其定位于配子体和卵囊壁上,为进一步解析EnOXIO1蛋白参与卵囊壁形成的分子机制奠定基础,为研制新型免疫阻断型球虫亚单位疫苗提供重要靶抗原。 相似文献
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根据已克隆的堆型艾美球虫(Eimeria acervulina)广东株子孢子表面抗原基因cSZ1的cDNA序列设计了特异性引物，用PCR方法扩增cSZ1的开放阅读框架(ORF)后克隆至表达载体pET-32a( )，构建了重组表达质粒pET-32a( )-cSZ1，并将其转化至大肠埃希氏菌BL21(DE3)。经IPTG诱导，获得了cSZ1重组抗原在大肠埃希氏菌中的高效表达，表达产物量可达菌体总蛋白的9．3％，融合蛋白的分子质量约为40ku。重组菌诱导表达的产物经SDS—PAGE后，用堆形艾美球虫感染鸡的超免疫血清进行免疫印迹分析，结果为阴性，提示cSZ1所编码的抗原可能主要是T细胞抗原表位。 相似文献
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The relationship between dry matter (DM) degradation and crude protein (CP) degradation in the dairy cow's rumen was determined with a view to defining the protein value of feeds for ruminants. The nylon bag technique was applied for these studies. For all the feeds investigate (green fodder and preserves from cocks-foot, ryegrass, alfalfa/grass and meadow grass, as well as alfalfa, extracted soybean meal) a significantly positive relationship was found to exist between the levels of DM and CP degradation (r = 0.73 to 1.0). The regression coefficient b1 (CP degradation as regressor) was found to average 0.87. The positive relationship between DM degradation and CP degradation implies that microbial protein amount and unfermented feed protein at the duodenum are negatively correlated. Model calculations show that, on account of the compensation between microbial protein and feed protein at the duodenum, in feeds with a CP concentration below 200 g/kg DM, the extent of ruminal protein degradation does not exert a marked influence on duodenal protein passage. The partial calculation of the duodenal protein supply on the basis of undegraded feed protein and microbial protein, as practiced in the new models of protein evaluation, leads to systematic errors unless the relationship between DM degradation and CP degradation is considered. 相似文献
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Spielmann J Kluge H Stangl GI Eder K 《Journal of animal physiology and animal nutrition》2009,93(4):400-409
This study was performed to assess the effects of potato protein and fish protein on concentrations of lipids in plasma and lipoproteins and the expression of genes involved in lipid metabolism in pigs used as an animal model. Therefore, 27 young male pigs with an average body weight of 22 kg were fed diets supplemented with protein extracted from potatoes (containing 849 g protein/kg dry matter), Alaska Pollack fillet as a source of fish protein (containing 926 g crude protein/kg dry matter) or casein which was used as control, for 3 weeks. Diets were formulated to supply identical amounts of each protein to the pigs by the three protein sources, namely 116 g/day in first week and 150 g/day in the second and third week. Pigs fed potato protein had lower concentrations of cholesterol in plasma and LDL than pigs fed casein (p < 0.05); no effect was observed on concentrations of HDL cholesterol and triglycerides. Pigs fed fish protein had lower cholesterol concentrations in plasma, LDL and HDL, and lower triglyceride concentrations in triglyceride-rich lipoproteins than pigs fed casein (p < 0.05). mRNA concentrations of genes involved in bile acid synthesis and cholesterol uptake were higher in pigs fed fish protein than in pigs fed casein (p < 0.05); no effect on these genes was observed in pigs fed potato protein. Expression of genes involved in lipogenesis and fatty acid oxidation was not altered by fish protein. In conclusion, this study shows that fish protein and potato protein lower plasma cholesterol concentrations in pigs. The hypocholesterolaemic effect of fish protein might be in part caused by a stimulation of bile acid synthesis; the reason for the hypocholesterolaemic effect of potato protein requires further elucidation. 相似文献
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1. Experiments were conducted to investigate whether or not varying dietary protein intake affects whole-body protein turnover rates in young chicks. 2. Seven-d-old single comb White Leghorn male chicks were fed on diets with protein concentrations of 0, 100, 200 or 400 g/kg diet under conditions of ad libitum or equalised feeding. At the end of the experiments, the rate of protein synthesis and protein degradation in the whole body were measured in vivo. 3. The results showed that both fractional and absolute rates of protein synthesis increased with increasing dietary protein up to 200 g/kg; above this concentration they remained almost constant when feeding was ad libitum. 4. Similar responses were found with equalized feeding except that a significant reduction in protein synthesis was found when dietary protein was increased from 200 to 400 g/kg diet. 5. Less sensitive and almost parallel changes in protein degradation rates were found. 6. It was concluded that adaptation to varied dietary protein intake occurred primarily through changes in protein synthesis, accompanied by parallel alterations in protein degradation in the whole body. 相似文献
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A. S. Green J. J. Ramsey C. Villaverde D. Asami A. J. Fascetti 《Journal of animal physiology and animal nutrition》2008,92(2):219-219
It is well-accepted that cats require more dietary protein than omnivores and herbivores. Work on hepatic enzyme activities showed that cats lack the ability to regulate the urea cycle enzymes in response to the dietary supply of protein. It was thus hypothesized that the high protein requirement of cats is due to an inability to regulate these enzymes, limiting adaptation to a low protein diet. We used indirect respiration calorimetry to assess the in vivo ability of cats to adapt substrate oxidation to different levels of dietary protein, including one below their protein requirement. In random order, eight cats consumed each of four semi-purified diets containing 7.7% (LP), 14.6% (AP), 27.3% (MP) and 51.1% (HP) of ME from protein. Cats consumed each diet for at least 14 days and then completed a 5-day nitrogen balance trial and at least 2, 12-hour indirect calorimetry measurements. The data were analyzed by anova using the Mixed procedure of SAS and are expressed as mean ± SEM. There was a significant effect of diet on protein oxidation (p < 0.0001), measuring 9.8 ± 0.5%, 13.4 ± 0.9%, 23.5 ± 0.8% and 49.0 ± 1.8% of total energy expenditure on the LP, AP, MP and HP diets, respectively. The ratio of protein oxidation/protein intake was significantly higher with the LP diet (1.27 ± 0.07) than the other three diets (AP, 0.92 ± 0.06; MP, 0.86 ± 0.03; HP, 0.96 ± 0.04; p < 0.0001), indicating a net loss of protein on the LP diet. Thus, cats adapted to a wide range of dietary protein concentrations, but were unable to fully adapt to the LP diet. 相似文献
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乳猪能否顺利断奶是养猪业成功的关键.乳猪消化系统的发育尚未完善,需要易消化、质量高的蛋白原料.因此,高质量、易消化的乳蛋白和血浆蛋白等常被用作乳猪开食料的蛋白原料. 相似文献