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猪α干扰素/白细胞介素2基因的融合表达及活性研究 总被引：5，自引：1，他引：4
为了研究高效广谱的猪基因工程抗病毒制剂,作者采用重叠延伸PCR(Splicing by overlap extension-PCR,SOE-PCR)方法通过一基因柔性接头(linker)(G4S)3将猪α干扰素(Porcine interferon alpha,PoIFN-α)与猪白细胞介素2(Porcine interleukin-2,PoIL-2)成熟肽基因连接,构建成PoIFN-α-linker-PoIL-2嵌合基因,并克隆入pGEM-T Easy载体,将PoIFN-α-linker-PoIL-2嵌合基因亚克隆入pQE-30表达载体进行原核表达.通过尿素变性、复性液复性、PBS溶液透析等步骤对表达的重组融合蛋白(rPoIFN-α-linker-PoIL-2)进行纯化.采用细胞病变抑制法检测rPoIFN-α-linker-PoIL-2蛋白中的PoIFN-α在不同细胞系上对不同病毒增殖活性的抑制作用;分别采用MTT法和PoIL-2 ELISA试验方法检测rPoIFN-α-linker-PoIL-2蛋白中PoIL-2的生物学活性.结果表明成功构建并克隆PoIFN-α-linker-PoIL-2嵌合基因.嵌合基因在大肠杆菌中得到高效表达,表达的rPoIFN-α-linker-PoIL-2蛋白相对分子质量约36.7 ku,蛋白经纯化后纯度在96%以上.rPoIFN-α-linker-PoIL-2蛋白在细胞上具有与单一rPoIFN-α蛋白相近的抑制病毒增殖活性,其中在PK-15细胞上抗VSV的活性单位为1.891×104 IU·mL-1,在Marc-145细胞上抑制高致病性PRRSV增殖的活性单位为905 U·mL-1;rPoIFN-α-linker-PoIL-2蛋白具有与单一rPoIL-2蛋白对照相近的生物学活性,可明显促进CTLL-2细胞的增殖,并可与抗PoIL-2单抗发生特异性免疫反应.这表明rPoIFN-α-linker-PoIL-2蛋白具有rPoIFN-α和rPoIL-2蛋白双重的生物学活性,为基因工程抗病毒制剂的开发以及高致病性猪繁殖与呼吸综合征等病毒性疾病的预防和治疗等研究奠定了基础. 相似文献
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猪α干扰素的原核表达及抗高致病性猪繁殖与呼吸综合征病毒活性测定 总被引：1，自引：0，他引：1
为了研究高效广谱的基因工程抗病毒制剂,本研究采用PCR技术自猪肝脏总DNA中扩增、克隆猪α干扰素（PoIFN-α）成熟肽基因并亚克隆入pQE30载体进行原核表达,对表达的融合重组猪α干扰素（rPoIFN-α）蛋白通过尿素变性、低浓度蛋白复性液复性、PBS溶液透析等步骤进行纯化,采用细胞病变抑制法分别测定rPoIFN-α蛋白在PK15细胞上抗水泡性口炎病毒（VSV）增殖活性及在Marc-145细胞上抑制高致病性猪繁殖与呼吸综合征病毒（PRRSV）的增殖活性,以分别评价rPoIFN-α的活性单位及其体外抑制高致病性PRRSV增殖所需的最低浓度。结果表明,克隆的PoIFN-α成熟肽基因全长501bp,编码166个氨基酸;表达的rPoIFN-α蛋白分子量大小20.7ku左右,与预期大小相一致;经纯化后的蛋白纯度在95%以上;纯化的rPoIFN-α蛋白在PK15细胞上抗VSV病毒活性单位为1.4482×10^4IU/mL（5.2×10^6IU/mg）,在Marc-145细胞上能完全抑制高致病性PRRSV增殖所需的rPoIFN-α蛋白最低有效剂量为640U/mL（2.3×10^4U/mg）。这些研究结果为新型基因工程抗病毒制剂的开发以及用于高致病性PRRSV性疾病的预防和治疗奠定了基础。 相似文献
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甘草酸体外抗PRRSV的作用机制 总被引：1，自引：0，他引：1
为了研究甘草酸体外抑制猪繁殖与呼吸综合征病毒(PRRSV)感染Marc-145细胞的作用及机制,本试验确定了甘草酸的抗PRRSV作用,并通过检测甘草酸对病毒的吸附侵入、核酸复制等环节对病毒的结构蛋白、非结构蛋白以及宿主细胞受体和IFN-α表达的影响确定甘草酸体外抗PRRSV的作用机制。结果显示,甘草酸阻断PRRSV感染时减少了GP4的表达,并降低CD163和CD151的表达;抑制PRRSV吸附时抑制了PRRSV GP4和细胞CD163的表达;抑制PRRSV侵入时抑制了GP4的表达,并显著提升细胞CD163和CD151的表达;抑制增殖的作用机制是抑制细胞内PRRSV nsp9和nsp10的表达,前期抑制病毒引起的TLR3表达升高的趋势,后期促进TLR3的表达;中后期降低、末期促进IFN-α的表达。结果表明,甘草酸能通过调节PRRSV的蛋白、宿主细胞受体和细胞因子的表达来抑制PRRSV体外感染Marc-145细胞。 相似文献
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1.大豆:包括黄豆和黑豆,生大豆及饼粕中含有3种有害物质,一是抗胰蛋白酶,它能抑制蛋白酶催化分解蛋白质;二是植物血球凝集素,能使血细胞减少、血红素含量降低;三是脲酶,会分解蛋白质和尿素生成氨,刺激消化道.这3种有害物质都可用加热、蒸煮、炒熟的办法进行脱毒. 相似文献
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乳蛋白是乳中重要的营养成分之一,超过90%的乳蛋白是乳腺利用从血液中摄取的氨基酸从头合成,因此在保证氨基酸充足供给的前提下,乳腺对氨基酸摄取率的高低是影响乳蛋白产量的关键因素。血液中的氨基酸不能自由扩散进出乳腺,需要由乳腺上皮细胞膜上特异的氨基酸转运载体(AAT)协助完成。而乳腺AAT活性受到营养物质和激素水平的调节,当乳腺感知到营养物质和激素水平变化的信号,能够通过激活或抑制以哺乳动物雷帕霉素靶蛋白复合物1(mTORC1)和一般性调控阻遏蛋白激酶2(GCN2)为核心的2条信号通路的活性,进而影响AAT活性,调节乳腺对氨基酸的摄取。本文主要从乳腺AAT的分类和功能、影响乳腺摄取氨基酸的主要因素以及调控乳腺氨基酸摄取的信号通路机制3个方面作一综述,旨在从氨基酸摄取的角度为提高乳蛋白的合成提供参考。 相似文献
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氨基酸转运载体（AAT）是一类介导氨基酸从细胞外转运到细胞内的重要蛋白，也是一类能介导氨基酸相关的信号通路的重要营养物质感受分子，在机体的生长代谢、营养健康等方面具有重要作用。动物机体中存在多种类型的AAT，它们能感知机体内相关氨基酸水平的变化，介导细胞氨基酸感知信号通路——哺乳动物雷帕霉素靶蛋白复合体1（mTORC1）和一般性调控阻遏蛋白激酶2（GCN2）的激活，从而引起通路下游发挥作用。在不同组织细胞中，发挥主导作用的AAT存在差异，表明AAT具有组织特异性，同时，AAT也受多种因素的影响，比如动物机体本身、营养物质水平、激素水平等。作者主要从AAT的类型及转运机制、介导营养信号启动及对mTORC1通路和GCN通路的影响、在不同组织中的作用及AAT表达的调控4个方面进行综述，从宏观方面介绍了AAT，旨在为AAT的研究提供一些参考。 相似文献
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A technique for the rapid field assessment of African animal trypanosomosis (AAT) was developed during studies in the Gambia. This involved gathering indigenous information from rapid-appraisal questionnaires addressed to local informants, the results of single tsetse surveys and evaluations of the prevalence of trypanosome infections in village cattle. Local informants included livestock owners and herdsmen and trained personnel such as livestock assistants. The answers to the questionnaires were weighted in order to translate them into semi-quantitative ranked estimates (zero, low, medium, high or very severe) of the severity of AAT problems. A similar ranking was also defined for tsetse and prevalence data in the Gambia. The three assessment methods generally gave complementary results leading to similar conclusions about the severity of tsetse-trypanosomosis problems in a survey area; inconsistencies usually suggested that additional information was needed. The rankings of AAT intensity were used to develop management guidelines for minimising the impact of AAT at different levels through control interventions or improved livestock management. The methodology was designed to provide reliable, up-to-date and cost-effective assessments of AAT problems. Emphasis was placed on the importance of the involvement, priorities and perceptions of village livestock owners and herdsmen in making these assessments. 相似文献
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Tzachi Zamir 《Society and Animals》2006,14(2):179-199
Is nonhuman animal-assisted therapy (AAT) a form of exploitation? After exploring possible moral vindications of AAT and after establishing a distinction between "use" and "exploitation," the essay distinguishes between forms of animal-assisted therapy that are morally unobjectionable and those modes of it that ought to be abolished. 相似文献
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基于GIS技术的青海省草地类型分类研究 总被引：6，自引：2，他引：4
分析青海省50个气象台站1971-2006年的＞0 ℃年积温平均值和年降水量平均值,以及各站的海拔高度和纬度值,建立年积温和年降水量的估算方程。在GIS环境下,根据年积温和年降水的估算方程,以及青海省高程资料,插值出分辨率为4 km×4 km的各个格点的年积温和年降水量值,然后根据湿润度公式计算出每个格点的湿润度值,最后根据综合顺序分类法第一级--类的检索图,确定各个格点的草地类型。研究结果表明,青海省天然草地共有18类,其中寒冷潮湿多雨冻原、高山草甸类的面积最大,约占草地总面积的50%。 相似文献
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摘要：肾阳虚证为命门火衰,温煦失常,气化不利所致的一种虚寒性证候,肾阳虚证在家畜生产实践中常见,影响其生产效能。治疗方剂层出不群,传统古方五子衍宗丸、补肾宁、肾气丸和右归丸等是常用基础方,在此基础上结合辨证施治方法衍生的自拟处方不在少数。对诸多方剂适用证候、治疗效果进行总结概括,指导兽医人员临床用药。结果表明：催情散多用于肾阳虚引起的公畜少精、弱精证,以及母畜卵巢发育不全等繁殖障碍性疾病;肾气丸与右归丸多用于改善公畜生殖系统,提高精子活力;补肾宁可促进母畜排卵,五子衍宗丸能抑制生殖系统炎症因子,降低生殖道炎症的发生率。 相似文献
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Livestock diseases have long been a challenge to livestock production and public health in sub-Saharan Africa and Zambia in particular. The Eastern Province of Zambia is one area in Zambia that is not spared by this challenge. Among various livestock diseases affecting cattle in this region, the most prominent are East Coast Fever (ECF) and African Animal Trypanasomiasis (AAT). Since little has been published on the epidemiological trends of these diseases in eastern Zambia, a retrospective epidemiological study was carried out using reports that were submitted to the provincial veterinary office over the past 20 years. This paper assists in evaluating the impact of some of these aid programmes. Data was analysed using Excel(?), SPSS(?), Epi Info(?), and Epi Map(?) software. Apparent prevalence of AAT in cattle had decreased in the study period from estimates as high as 50% in Katete and Petauke district in 1990 and 1992 respectively to just below 3% (Petauke and Katete) in 2008, thereby, reducing the provincial apparent prevalence from 20% in 1992 to just below 3% in 2008. AAT apparent prevalence dropped from estimates as high as 17% in Chadiza district and 6% in Chipata district in 1990 to just below 1% in 2008 thereby reducing the provincial mean prevalence of East Coast Fever from 6% (1990) to 1% (2008). The inclusion of donor assistance in disease control programmes for both AAT and ECF appeared to have a significant impact on the prevalence of both diseases. 相似文献
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Wangoola Robert Mandela Kevin Bardosh Acup Christine Among Welburn Susan Waiswa Charles Bugeza James 《Tropical animal health and production》2019,51(7):2011-2018
African animal trypanosomiasis (AAT) continues to inflict heavy losses on livestock production especially cattle in terms of decreased production and productivity in Uganda. AAT is a disease complex caused by tsetse fly-transmitted Trypanosoma brucei brucei, Trypanosoma brucei rhodesiense, Trypanosoma congolense, and Trypanosoma vivax. The disease is most important in cattle but also known to cause serious losses in pigs, camels, goats, and sheep. Several control measures including live bait technology, mass treatment of cattle with trypanocidal drugs, and deployment of tsetse traps have been used in the past 10 years, but the problem still persists in some areas. This necessitated an exploration of the factors associated with continued trypanosome infections in cattle, which are also known reservoirs for the zoonotic trypanosomiasis. A structured questionnaire was administered to 286 animal owners from 20 villages purposively selected from Lira, Kole, and Alebtong districts of Lango subregion to obtain information on the factors associated with persistence of infection. Over 50% of the respondents reported trypanosomiasis as a major challenge to their livestock. Land ownership (P = 0.029), type of livestock kept (P = 0.000), disease control strategy employed (P = 0.000), source of drugs (P = 0.046), and drug preparation (P = 0.017) were associated with persistent AAT infection. We recommend continued farmer sensitization on the threat of AAT and the available prevention and control options. The use of isometamidium chloride for prophylaxis against trypanosomiasis is highly recommended. There is also a need to foster qualified private veterinary drug supply in the region.
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Waltner-Toews D 《The Canadian veterinary journal. La revue veterinaire canadienne》1993,34(9):549-551
A survey was done of 150 systematically selected United States animal care agencies and 74 Canadian humane societies to determine the prevalence of animal assisted therapy (AAT) programs; concerns about, and experience with, zoonotic diseases; and precautions taken to prevent zoonotic disease transmission. Of the 69 US agencies and 49 Canadian societies that reported having AAT programs, 94% used dogs and/or cats in their programs, 28% used rabbits, 15% used “pocket pets” (hamsters, gerbils, mice, guinea pigs), and 10% used birds (excluding poultry). About two-thirds of the programs were involved with the elderly in nursing homes, about a quarter of them worked with schools, and a quarter worked with hospitals. Half of the respondents had concerns about zoonotic disease control. Rabies, ringworm, and external parasitism were the most commonly cited zoonotic diseases of concern. Few concerns were based on actual experience. Fewer than half of the programs consulted a health professional about prevention of zoonotic diseases. Only 10% of the respondents reported having printed guidelines about the prevention of zoonotic disease transmission. Practising veterinarians are encouraged to make their expertise available to local AAT programs. 相似文献
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Non-cytopathogenic bovine viral diarrhoea virus (ncpBVDV) has previously been shown to inhibit the function of interferon regulatory factor-3 in cultured cells [J. Virol. 76 (2002) 8979]. In this study, we show that, like ncpBVDV, when cells were previously exposed to cytopathogenic BVDV (cpBVDV) the appearance of an IRF-3–DNA complex from nuclear extracts that can be induced by heterologous virus infection was not observed. Infection of cells with ncpBVDV or cpBVDV resulted in neither the translocation of IRF-7 from the cytoplasm to the nucleus of infected cells, nor an inhibition of its nuclear translocation in cells super-infected by Semliki Forest Virus. We conclude that cpBVDV and ncpBVDV both share the ability to inhibit the full function of IRF-3 but neither stimulate or block the nuclear uptake of IRF-7. 相似文献