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Impact of an oligosaccharide-based polymer on the metabolic profiles and microbial ecology of weanling pigs experimentally infected with a pathogenic E.coli
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作者 Kwangwook Kim Cynthia Jinno +4 位作者 Xunde Li David Bravo Eric Cox Peng Ji Yanhong Liu 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2024年第2期749-764,共16页
Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to ... Background Our previous study has reported that supplementation of oligosaccharide-based polymer enhances gut health and disease resistance of pigs infected with enterotoxigenic E.coli(ETEC)F18 in a manner similar to carbadox.The objective of this study was to investigate the impacts of oligosaccharide-based polymer or antibiotic on the host metabolic profiles and colon microbiota of weaned pigs experimentally infected with ETEC F18.Results Multivariate analysis highlighted the differences in the metabolic profiles of serum and colon digesta which were predominantly found between pigs supplemented with oligosaccharide-based polymer and antibiotic.The relative abundance of metabolic markers of immune responses and nutrient metabolisms,such as amino acids and carbohydrates,were significantly differentiated between the oligosaccharide-based polymer and antibiotic groups(q<0.2 and fold change>2.0).In addition,pigs in antibiotic had a reduced(P<0.05)relative abundance of Lachnospiraceae and Lactobacillaceae,whereas had greater(P<0.05)Clostridiaceae and Streptococcaceae in the colon digesta on d 11 post-inoculation(PI)compared with d 5 PI.Conclusions The impact of oligosaccharide-based polymer on the metabolic and microbial profiles of pigs is not fully understood,and further exploration is needed.However,current research suggest that various mechanisms are involved in the enhanced disease resistance and performance in ETEC-challenged pigs by supplementing this polymer. 展开更多
关键词 CARBADOX Colon microbiota Enterotoxigenic e.coli f18 Metabolomics Oligosaccharide-based polymer Weaned pigs
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F18^+E.coli株可引起生长-肥育猪腹泻
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作者 金福源 陶艳华 《国外畜牧学(猪与禽)》 2011年第4期42-45,共4页
大肠杆菌是引起断奶仔猪腹泻的一种常见致病因子,但通常不会影响生长-肥育猪。最初认为11周龄生长-肥育猪出现的急性、严重水样腹泻是传染性胃肠炎。在对送检样本进行检测后表明,致病因子是能够产生志贺样毒素的F18^+E.coli。患病猪无... 大肠杆菌是引起断奶仔猪腹泻的一种常见致病因子,但通常不会影响生长-肥育猪。最初认为11周龄生长-肥育猪出现的急性、严重水样腹泻是传染性胃肠炎。在对送检样本进行检测后表明,致病因子是能够产生志贺样毒素的F18^+E.coli。患病猪无神经症状或其他的与水肿病有关的症状,新霉素治疗有效。临床受感染猪群与未受感染猪群在死亡率和生长性能上无明显的差异。本文报告了11周龄生长-肥育猪感染F18^+E.coli株的罕见病例。 展开更多
关键词 腹泻 大肠杆菌(e.coli) f18 志贺样毒素
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猪miR-192/-215的组织表达谱及其关键靶基因分析 被引量:6
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作者 吴正常 殷学梅 +4 位作者 孙丽 夏日炜 霍永久 吴圣龙 包文斌 《中国农业科学》 CAS CSCD 北大核心 2015年第11期2251-2261,共11页
【目的】在前期通过Illumina Solexa高通量测序技术并结合荧光定量验证筛选出断奶仔猪E.coliF18菌株感染下表达量极显著上调的micro RNAs—miR-192和miR-215的基础上,为了解miR-192和miR-215的组织表达情况,进一步筛选确定miR-192和miR-... 【目的】在前期通过Illumina Solexa高通量测序技术并结合荧光定量验证筛选出断奶仔猪E.coliF18菌株感染下表达量极显著上调的micro RNAs—miR-192和miR-215的基础上,为了解miR-192和miR-215的组织表达情况,进一步筛选确定miR-192和miR-215靶向调控E.coli F18菌株感染的关键靶基因,为探讨miR-192和miR-215调控断奶仔猪E.coli F18感染的分子调控机制奠定基础。【方法】试验采用Real-time PCR方法检测miR-192和miR-215在35日龄梅山仔猪各个组织中的分布情况,同时利用MEGA 5.0分析了其保守性,Target Scan预测miR-192和miR-215的靶基因,并对靶基因分别进行Gene Ontology和Pathway通路的富集分析,进一步利用DAVID对靶基因蛋白进行功能分类,最后将预测的靶基因与课题组前期筛选出的断奶仔猪E.coli F18感染抗性相关的调控基因取交集。【结果】miR-192和miR-215在35日龄梅山仔猪十二指肠和空肠中均高度表达,二者成熟序列在脊椎动物中高度保守。miR-192和miR-215对应的靶基因相同,具有156个靶基因,只在轴突导向通路(axon guidance pathway)显著富集(P-value<0.05)以及25个GO功能中显著富集(P-value<0.05)。靶基因蛋白按功能分为12类,其中信号转导功能的磷蛋白(phosphoprotein)和DNA结合蛋白(dna-binding)占主要部分。靶基因与前期筛选的E.coli F18感染抗性相关的调控基因(GEO登录号:GSE26854)取交集,发现DLG5、ALCAM、FRMD4B、MIPOL1和ZFHX3等5个基因为miR-192和miR-215的重要靶基因,其中DLG5为维持小肠上皮细胞结构完整性的重要因子。【结论】miR-192与miR-215是参与维持断奶仔猪肠道正常功能与抗F18大肠杆菌感染的重要因子,DLG5可能是miR-192和miR-215靶向调控E.coli F18菌株感染中关键的靶基因,今后需要进一步验证其能否作为梅山猪抗F18大肠杆菌的有效遗传标记。 展开更多
关键词 miR-192 miR-215 组织表达谱 靶基因 e.coli f18
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大鼠肝脏F蛋白基因的克隆和表达
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作者 刘树业 俞新大 +3 位作者 宋春娟 张健东 石欣荣 张琚 《同济大学学报(医学版)》 CAS 2007年第1期10-14,共5页
目的获得大鼠肝脏F蛋白基因克隆(F-protein’s cDNA);利用原核(大肠杆菌)表达系统表达大鼠肝脏F蛋白。方法提取大鼠肝脏总RNA,对其进行反转录和PCR(RT-PCR)扩增出目的基因(F-protein’s cDNA),然后将其与pUCm-T载体进行连接获得克隆质粒... 目的获得大鼠肝脏F蛋白基因克隆(F-protein’s cDNA);利用原核(大肠杆菌)表达系统表达大鼠肝脏F蛋白。方法提取大鼠肝脏总RNA,对其进行反转录和PCR(RT-PCR)扩增出目的基因(F-protein’s cDNA),然后将其与pUCm-T载体进行连接获得克隆质粒pUCm-T-F并转化到大肠杆菌DH5α中;将测序结果正确的克隆片断重新与表达载体pET-15b连接,获得F抗原表达质粒pET15b-F并将其转化到表达菌株BL21(DE3)pLysS中,用1mmol/L IPTG诱导其表达。结果RT-PCR扩增出的目的基因(F-protein’s cDNA),经测序证明与Gene-bank上提供的F蛋白cDNA序列完全一致;表达后的全菌蛋白进行SDS-PAGE电泳检测,目的蛋白分子量大小约为43 kD,与预期值相符,表达量可达全菌总蛋白量的40%。结论表达的目的蛋白经His-tag柱进行亲和层析纯化,SDS-PAGE检测得到了不含其他杂蛋白的单一F蛋白条带,与豚鼠抗人F蛋白抗血清反应成阳性,说明我们经基因工程方法得到的纯化的F蛋白有免疫活性。 展开更多
关键词 肝脏 F抗原 大肠杆菌 基因克隆 蛋白表达 大鼠
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人乳头瘤病毒18型L1蛋白在大肠埃希菌中的可溶性表达及病毒样颗粒的形成
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作者 安静 付生芳 +4 位作者 李雄雄 包红 寇桂英 白幕群 余黎 《微生物学免疫学进展》 2014年第5期11-16,共6页
目的利用大肠埃希菌系统可溶性表达人乳头瘤病毒18型(HPV18)L1蛋白,纯化和重组装获得HPV18病毒样颗粒(VLPs),为进一步研制HPV18基因工程疫苗奠定基础。方法首先按大肠埃希菌密码子偏好进行HPV18L1全基因合成,经PCR扩增出截短的HPV18L1基... 目的利用大肠埃希菌系统可溶性表达人乳头瘤病毒18型(HPV18)L1蛋白,纯化和重组装获得HPV18病毒样颗粒(VLPs),为进一步研制HPV18基因工程疫苗奠定基础。方法首先按大肠埃希菌密码子偏好进行HPV18L1全基因合成,经PCR扩增出截短的HPV18L1基因,构建重组表达载体PET30a-L1,通过优化表达在大肠埃希菌BL21中可溶性表达L1蛋白,其次采用硫酸铵沉淀、离子交换层析、疏水层析后,获得高纯度的的L1蛋白,再通过解聚和重聚获得VLPs。结果全基因优化并截短的HPV18L1蛋白在大肠埃希菌系统中以可溶形式表达,纯化后的蛋白纯度达到90%以上,电镜下观察到直径为60 nm的VLPs颗粒。结论利用大肠埃希菌系统可溶性表达非融合HPV18L1蛋白,并获得均一的VLPs颗粒,为疫苗的开发奠定基础。 展开更多
关键词 大肠埃希菌表达系统 人乳头瘤病毒18型 L1蛋白 优化 可溶性表达 病毒样颗粒
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不同表达载体对人白细胞介素-18原核表达效率的影响 被引量:4
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作者 吉莉莉 杨吉成 +2 位作者 孙万邦 盛伟华 李丽蛾 《遵义医学院学报》 2003年第1期4-6,共3页
目的 探讨PBV220及PJW2载体构建的rhIL-18重组质粒的IL-18表达效率。方法 将构建的PBV220/rhIL-18重组质粒中的rhIL-18酶切、克隆到表达载体PJW2中,经PCR产物电泳及测序鉴定,热诱导后在大肠杆菌中表达。结果 PJW2/rhIL-18表达的重组人IL... 目的 探讨PBV220及PJW2载体构建的rhIL-18重组质粒的IL-18表达效率。方法 将构建的PBV220/rhIL-18重组质粒中的rhIL-18酶切、克隆到表达载体PJW2中,经PCR产物电泳及测序鉴定,热诱导后在大肠杆菌中表达。结果 PJW2/rhIL-18表达的重组人IL-18占菌体总蛋白质的20%,而PBV220/IL-18重组质粒表达的重组人IL-18占菌体总蛋白质的16%。两者表达的蛋白质分子量约为18KD,与预期分子量相符。结论 PJW2载体rhIL-18表达效率高于PBV220,为下一步的纯化及提高蛋白产量打下了基础。 展开更多
关键词 白细胞介素18 表达 载体 大肠杆菌
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志贺氏菌样毒素Stx2e缺失突变株的构建及部分生物学特性的分析
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作者 张志强 姜露 +2 位作者 李永慧 段强德 朱国强 《中国动物传染病学报》 CAS 2013年第1期53-59,共7页
水肿病是能由产志贺样毒素型大肠杆菌产生的Stx2e毒素引起的一种细菌传染病,对断奶仔猪有较高的致死率,常造成较大的经济。当前,Stx2e毒素影响病原菌与靶细胞之间的相互作用机制不明确,Stx2e基因缺失株作为后选疫苗的潜力有待确认。本... 水肿病是能由产志贺样毒素型大肠杆菌产生的Stx2e毒素引起的一种细菌传染病,对断奶仔猪有较高的致死率,常造成较大的经济。当前,Stx2e毒素影响病原菌与靶细胞之间的相互作用机制不明确,Stx2e基因缺失株作为后选疫苗的潜力有待确认。本研究中我们利用Red重组系统构建了大肠杆菌1522(serotype O139:K12:H1)和1525(serotype O157:H19)的Stx2e基因缺失株,并通过PCR检验,DNA测序手段以及Vero细胞毒性试验在细胞水平加以验证。研究发现,在同样的培养条件下,1522△Stx2e株的生长特性与原型株相比没有差异,其分解发酵糖类和氨基酸的能力也未发生改变,但是相较野生株,Stx2e基因缺失株粘附猪肠上皮细胞IPEC-J2的能力大幅度降低,可达30%。将表达K99菌毛操纵子基因克隆入1522△Stx2e中,该重组菌能够同时表达K99菌毛和自身的F18菌毛基因,且该重组株的粘附IPEC-J2细胞能力相较原型株增强近1倍。 展开更多
关键词 大肠杆菌 f18菌毛 RED重组系统 Stx2e基因 突变株
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Zinc phosphate-based nanoparticles as alternatives to zinc oxide in diet of weaned piglets 被引量:1
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作者 Silvia Kociova Kristyna Dolezelikova +14 位作者 Pavel Horky Sylvie Skalickova Daria Baholet Lucie Bozdechova Eva Vaclavkova Jaroslava Belkova Pavel Nevrkla Jiri Skladanka Tomas Do Ondrej Zitka Yazan Haddad Pavel Kopel Ludek Zurek Vojtech Adam Kristyna Smerkova 《Journal of Animal Science and Biotechnology》 SCIE CAS CSCD 2020年第4期1147-1162,共16页
Background: The high doses of zinc oxide(Zn O) administered orally to piglets for the prevention of diarrhea and increase of growth rate can contaminate pig farms and the surrounding environment. Therefore, there is a... Background: The high doses of zinc oxide(Zn O) administered orally to piglets for the prevention of diarrhea and increase of growth rate can contaminate pig farms and the surrounding environment. Therefore, there is a need to find a replacement of high doses of dietary Zn O with an equally effective alternative. In the present study, the effect of two formulations of zinc phosphate-based nanoparticles(Zn A and Zn C NPs) on growth performance,intestinal microbiota, antioxidant status, and intestinal and liver morphology was evaluated. A total of 100 weaned piglets were randomly divided into 10 equal groups with the base diet(control) or the base diet supplemented with Zn A, Zn C, or Zn O at concentrations 500, 1000, and 2000 mg Zn per kilogram of diet. Supplements were given to animals for 10 days. Fecal samples were collected on day 0, 5, 10 and 20. At the end of the treatment(day 10),three piglets from each group were sacrificed and analyzed.Results: Comparing to that of control, the significantly higher piglet weight gain was observed in all piglet groups fed with Zn A(P < 0.05). Differences in the total aerobic bacteria and coliform counts in piglet feces after NPs supplementation compared to that of control and Zn O groups were also found(P < 0.05). The majority of aerobic culturable bacteria from the feces represented Escherichia(28.57–47.62%), Enterococcus(3.85–35.71%), and Streptococcus(3.70–42.31%) spp. A total of 542 Escherichia coli isolates were screened for the virulence genes STa,STb, Stx2, F4, and F18. The substantial occurrence of E. coli virulence factors was found on day 5, mainly in fimbrillary antigen and thermostable toxins, except for piglets fed by Zn C. Zn treatment decreased Zn blood levels in piglets fed with Zn O and Zn A(500 mg/kg) and increased in Zn C(2000 mg/kg) compared to that of control(P < 0.05). The antioxidant status of piglets was affected only by Zn A. While some changes in the liver and the intestinal morphology of piglets with NPs were observed, none were serious as reflected by the normal health status and increased weigh gain performance.Conclusions: Our results indicate that Zn A NPs have a positive effect on the piglet growth performance even at the lowest concentration. The prevalence of E. coli virulence factors was lowest in pigs supplemented with Zn C.Zinc phosphate-based nanoparticles may be an effective alternative to Zn O. 展开更多
关键词 Antioxidant status DIET e.coli STa STB Stx2 F4 f18 Fecal coliforms MICROBIOTA
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Development of an Indirect ELISA for Detection of T.solium Based on Recombinant 18 kDa Protein
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作者 WU Guo-hua ZHENG Ya-dong +5 位作者 JIA Wan-zhong ZHANG Shao-hua JING Zhi-zhong LUO Xue-nong LIU Shi-quan CAI Xue-peng 《畜牧兽医学报》 CAS CSCD 北大核心 2010年第S1期87-92,共6页
The gene encoding the 18 kDa protein of Taenia solium metacestodes was amplified by RT-PCR and cloned into the pGEM-T vector for sequencing.The recombinant plasmid named pGEX-CE18 was constructed and transformed into ... The gene encoding the 18 kDa protein of Taenia solium metacestodes was amplified by RT-PCR and cloned into the pGEM-T vector for sequencing.The recombinant plasmid named pGEX-CE18 was constructed and transformed into E.coli BL21 for in vitro expression.SDS-PAGE and Western blot were employed for analyzing the recombinant protein,which was then used for development of an indirect ELISA for detection of anti-cysticercosis antibodies.The results showed that the recombinant protein of interest was 35 kDa in size,accounting for 28%of total bacteria proteins,and reacted with positive sera against cysticercosis.Using the newly-constructed indirect ELISA and a commercially available ELISA kit,paired analyses of 178 serum samples indicated that the concordant rate was 98.83%and the ELISA exhibited good specificity and sensitivity,supporting its utility and application for diagnosis of cysticercosis. 展开更多
关键词 CYSTICERCUS cellulosae 18 kDa PROTEIN e.coli EXPRESSION RECOMBINANT PROTEIN ELISA
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