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禽痘病毒载体及其应用 总被引：1，自引：0，他引：1
禽痘病毒(fowlpox virus,FPV)载体在近几年受到越来越多学者的关注。禽痘病毒载体的原理,是把外源基因插入到病毒的适当位置,通过同源重组把外源基因重组到病毒基因组的复制非必需区域。外源基因通过修饰等过程可以得到高效的表达,表达产物生物活性及相关功能没有改变,刺激机体产生较强的免疫应答。禽痘病毒被用作表达载体后,相继成功地表达了来源于禽类病毒、动物病毒,乃至能感染人类病原体的保护性抗原基因,现已被广泛用作基因工程的重要工具。作者就禽痘病毒粒子的结构、生物学、分子生物学、禽痘病毒载体及其应用等几个方面进行了阐述。 相似文献
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禽痘病毒表达载体是已构建表达外源基因的病毒载体中的一种，它在表达外源基因上具有明显的优势：(1)它的基因组结构庞大，能耐受较大的外源基因(25～35kb)在其基因组的不同位点插入。(2)严格的胞浆内复制，避免了病毒基因重组入宿主细胞染色体的可能性。且禽痘病毒的宿主范围较窄，仅感染禽类，在哺乳动物体内仅产生一过性感染，安全性较高。(3)表达产物(蛋白质)在翻译后的加工修饰过程与体内极为相似，因此活性高。且保留了相应的抗原性、免疫原性。基于这些特点，使得禽痘病毒载体不仅可作为禽源疫苗，而且还可作为哺乳动物乃至人类的基因工程活载体疫苗，具有广阔的应用前景。 相似文献
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禽痘病毒载体疫苗研究进展 总被引：1，自引：0，他引：1
禽痘病毒和其它痘病毒一样,基因组的高保守区的非复制域可以整合外源DNA,已成为普遍的表达载体,主要应用于痘病毒分子生物学领域、体外生产和蛋白质功能化研究以及作为活的疫苗或工具用于疫苗研究.禽痘病毒载体疫苗不仅能预防禽痘的发生,而且通过插入多个其它病原的保护性抗原基因以诱导产生抗外来抗原的抗体和细胞毒性T细胞反应,提高对相应病原的免疫抵抗力.由于重组病毒所表达的外源基因在机体内随载体病毒的复制而表达,与灭活疫苗相比,它容易生产,免疫剂量小,接种方法简便,接种一次就能获得长期的免疫效果,大大降低了生产成本. 相似文献
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以GPT和GFP为双重筛选标记的禽痘病毒转移载体的构建及鉴定 总被引：1，自引：0，他引：1
为研制和开发以禽痘病毒为载体的重组病毒疫苗,本研究构建了禽痘病毒通用转移载体pSY681-gfp-gpt,该载体含有gfp和gpt2种报告基因及背对的痘苗病毒晚期启动子P11和早期启动子P7.5,P11启动gfp和gpt两个基因,P7.5用于启动外源基因,在早期启动子P7.5下游引入NotⅡ和AftⅡ两个酶切位点.用于外源基因的插入.为检测禽痘病毒转移载体pSY681-gfp-gpt的有效性,将H9亚型禽流感病毒A/Chicken/Shanghai/10/01(H9N2)的HA基因插入到该载体中构建转移载体pSY681-gfp-gpt-HA9,将转移载体转染已感染禽痘病毒S-FPV-017的鸡胚成纤维细胞,利用gfp和gpt同时进行双重筛选、数轮蚀斑纯化后获得了重组病毒rFPV-gpt-gfp-HA9,通过PCR、western blot鉴定,结果证明,获得了能稳定表达H9亚型禽流感病毒HA蛋白的重组禽痘病毒rFPV-gpt-gfp-HA9,为今后重组禽痘病毒活载体疫苗的研制奠定了基础. 相似文献
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禽痘病毒的分子生物学Ⅱ禽痘病毒载体（续）王志亮综述刘秀梵审校（扬州大学农学院动物医学系）２．３外源基因痘病毒转录的特点决定了外源基因必须符合以下原则：２．３．１外源基因应是无自身启动子的单纯编码阅读框，因为痘病毒只能识别自家的启动子。２．３．２外源... 相似文献
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《中国预防兽医学报》2018,(12)
为研究表达鹅γ干扰素(GoIFN-γ)基因重组禽痘病毒的抗病毒活性,本研究构建了在禽痘病毒(FPV)早晚期启动子LP2EP2控制下的IFN-γ基因重组禽痘病毒转移载体,将其转染至亲本禽痘病毒S-FPV-017预感染的鸡胚成纤维细胞(Chicken embryo fibroblasts,CEF),使其与禽痘病毒基因组进行同源重组,经过9轮筛选和纯化并进行PCR和间接免疫荧光检测,获得能稳定表达外源基因的重组病毒r FPV-IFNγ-LacZ。利用细胞病变抑制法检测抗病毒活性结果显示,在鸡胚成纤维细胞中重组禽痘病毒表达的IFN-γ对鹅细小病毒的复制具有显著抑制作用。本研究为制备共表达保护性抗原和γ干扰素基因的重组基因工程疫苗研究奠定基础。 相似文献
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1 鸡痘病毒的特点、活载体的构建及作为活载体的优越性1.1 重组鸡痘病毒的特点及活载体的构建 鸡痘病毒(Fowlpox Virus,FPV)属于痘病毒科禽痘病毒属,含很大的双链DNA分子约300kb,其中存在许多病毒本身复制的非必需区(nonessential region,NER),并预计可容纳远不止25 kb的外源基因。这就可以用来研究和开发基因 相似文献
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Avipoxvirus is known to be endemic in New Zealand and it is a cause of ongoing mortalities in the endangered black robin and shore plover populations. There is no information on the strains of avipoxvirus occurring in New Zealand and their likely origin or pathogenicity. This study was designed to identify the phylogenetic relationships of pathogenic avipoxvirus strains infecting introduced, native, and endemic bird species in New Zealand. Avipoxvirus 4b core protein gene was detected in tissue samples from 25/48 birds (52.1%) from 15 different species in New Zealand. Bootstrap analysis of avipoxvirus 4b core protein gene revealed that the New Zealand avipoxvirus isolates comprised of three different subclades. The majority of New Zealand avipoxvirus isolates (74%) belonged to A1 subclade which shared 100% genetic similarity with the fowlpox HPB strain. An isolate from a wood-pigeon (kereru) belonged to subclade A3, displaying 100% sequence homology to albatrosspox virus. An additional group, isolated from two shore plovers and one South Island saddleback, grouped within subclade B1 and presented 99% sequence homology to European PM33/2007 and Hawaiian HAAM 22.10H8 isolates. The results suggest that a variety of New Zealand bird species are susceptible to avipoxvirus infection, that there are more than two distinctive avipoxvirus subclades in New Zealand, and that the most prevalent A1 strain may have been introduced to New Zealand through introduced avian hosts such as passerines or poultry. 相似文献
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Four great tits (Parus major) with nodular lesions suggestive of poxvirus infections were observed in a garden in Vienna, Austria. One of the birds was submitted for examination. Because of its poor condition, the bird had to be euthanatized and was subsequently necropsied. An avipoxvirus infection was confirmed by histopathology, electron microscopy, and polymerase chain reaction and sequencing. This is the second report of naturally occurring poxvirus infection in great tits and the first of its kind in central Europe. 相似文献
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Tarello W 《Veterinary dermatology》2008,19(2):101-104
The prevalence, cutaneous manifestations and concurrent clinical signs of avipoxvirus infection in 3706 falcons in two Middle Eastern countries are reported. Diagnosis was based on evidence of typical 'dry' skin lesions on featherless parts of the body and microscopic detection of Bollinger bodies in epithelial cells. Avipoxvirus was isolated from one representative case. Overall prevalence of cutaneous changes due to avipoxvirus infection did not differ significantly between Kuwait (2.7%) and Dubai (2.3%), although pox lesions were more prevalent on the feet of birds from Kuwait (67.4% vs. 50%) and more common on the eyelids of birds from Dubai (45.6% vs. 30.4%). Foot lesions were always present in birds with multiple infection sites. Some birds from Dubai had severe infection associated with weight loss, anorexia, lethargy, vomiting, central nervous system involvement and ultimately death. 相似文献
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为建立检测鸭痘病毒的TaqMan荧光定量PCR方法,本试验克隆了鸭痘病毒P4b基因,构建重组质粒pMD-DPV-P4b,并将其作为标准阳性模板。参照GenBank收录的禽痘病毒P4b基因设计合成1对特异性引物及与该引物相匹配的特异探针。以定量的10倍系列稀释的质粒pMD-DPV-P4b为标准品,通过对反应条件进行优化,建立了一种检测鸭痘病毒的TaqMan荧光定量PCR方法。结果显示,该方法与禽流感病毒、鸭黄病毒、鸭肝炎病毒、新城疫病毒、鸭瘟病毒和小鹅瘟病毒等其他水禽病毒,以及山羊痘病毒和鸡痘病毒等其他痘病毒均无交叉反应,特异性好。该方法最低检测限为1.29×102拷贝/μL,比普通PCR检测方法高100倍。组内和组间变异系数均小于2%。结果表明,本试验所建立方法具有灵敏、特异、安全、快速的特点,适用于鸭痘病毒的检测。 相似文献
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CAO Hui-hui SUN Wen-chao ZHENG Min WEI Xian-kai SU Jiao-xiu LIANG Sheng ZHENG Lie-feng LI Jun LIU Qi 《中国畜牧兽医》2015,42(10):2560-2566
To establishment a TaqMan Real-time PCR method for detection of duck poxvirus (DPV),we cloned the P4b gene of DPV.The specific primers and probe were designed according to the nucleotide sequence of avipoxvirus available in GenBank.Recombinant plasmid pMD-DPV-P4b was employed as positive standard template for Real-time PCR.By optimization of reaction conditions,a TaqMan Real-time PCR method for detection of DPV was established.The results of specificity test proved this method had no cross-react with other waterfowl vial agents and poxviruses including avian influenza virus,duck flavivirus,duck hepatitis virus,Newcastle disease virus,duck entertitis virus,goose parvovirus,goatpox virus and fowlpox virus.The detection limit of the assay was 1.29×102 copies/μL of viral DNA,which was 100 times higher than that of the routine PCR.Reproducibility test showed that the CVs of intra assay and inter assay were both less than 2%.Above results supported that the assay was suitable for the detection of DPV very well. 相似文献
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The ultrasonographic findings of fourteen animals having gastrointestinal foreign bodies are presented. Ultrasonography confirmed the presence and location of foreign material within the gastrointestinal tract when radiographic findings were unclear. The acoustic patterns arising from each foreign body varied depending on its physical properties and interaction with the ultrasound beam. These patterns, some of which seemed characteristic for certain types of foreign bodies, and the associated findings of mechanical ileus, ascites, mesenteric lymph node enlargement and localized pancreatitis are discussed. 相似文献
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武芳 《四川畜牧兽医学院学报》2008,(1):189-191
随着现代教育技术的不断发展,多媒体被引入了外语语言教学,它将计算机技术与语言教学结合起来,不但可以加强学生对外语语言类课程的理解,还能培养学生在实际语言应用环境中的实践能力。可以说,多媒体是连接学生语言知识和语言运用能力的一座桥梁。文中就多媒体环境下进行外语语言教学所具有的优势、应具备的条件以及存在的问题作了一些探讨。 相似文献
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Beatriz Moreno‐Aguado Ines Carrera Andrew Holdsworth Petra Agthe Thomas W. Maddox Tim Trevail 《Veterinary radiology & ultrasound》2020,61(4):417-426
The diagnosis and treatment of nasal foreign bodies usually includes a combination of rhinoscopy and imaging techniques, such as CT. The purpose of this retrospective, multicenter study was to describe the CT characteristics of nasal foreign bodies in dogs and cats and to determine if different nasal CT features exist between acute and chronic cases. Twenty dogs and six cats met the inclusion criteria. Eleven nasal foreign bodies (42%) were detected confidently with CT. The foreign body had a linear shape in 81% of cases and displayed a “tubular‐like appearance” in 54% of cases. In five cases (19%), a foreign body was suspected but not clearly visible. Additional CT changes were present in the nasal passages in 96% of the cases. The presence of turbinate destruction (P = .021) and mucosal thickening (P = .014) on CT were associated with the presence of a chronic nasal foreign body. In this sample, the nature of the foreign body did not influence its visibility and was not associated with specific CT characteristics. Computed tomography may be useful in the investigation of nasal foreign bodies, however, a negative CT examination does not exclude their presence. 相似文献
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Zahid R. Shah MD Jeffrey R. Crass MD Dubravka C. Oravec MD Errol M. Bellon MD 《Veterinary radiology & ultrasound》1992,33(2):94-100
Foreign bodies in the soft tissues are frequent problems in both veterinary and human medical practice. Radiography has been the principle tool for diagnosis, but is unable to define many foreign bodies. This study was undertaken to compare the sonographic and radiographic visualization for a variety of foreign bodies. Fresh turkey muscle (pectoral and thigh) with skin was used as the soft tissue model. Assorted foreign bodies were introduced 1 cm into the muscle. Materials examined were small wood fragments, steel wire, glass, nails, graphite, rock, BB pellets, and polystyrene plastic. Following introduction of the foreign bodies radiographs were obtained and the specimens were studied sonographically at 5.0 mHz and 7.5 mHz. All foreign bodies were visible with sonography. A bright echo with either acoustic shadowing or reverberations was present for all specimens. Radiographically metallic foreign bodies were visible but wood, some graphite, and plastic were not seen. We conclude that radiography should remain the primary imaging modality for foreign body detection because of its general availability and low cost. If radiography is negative ultrasound can detect and guide the removal of radiolucent foreign bodies. 相似文献
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草业科技英语具有专业性强、科技性强等特点,译者在翻译时很难把握,不能有效地传播和推广国外先进的草业科学技术。举例阐述了草业科技英语词汇及语法特点,从而提出了词汇、句子以及语篇的翻译技巧,有助于草业工作者进行文献资料的英汉翻译,从而进一步推进草业科技的推广和信息的交流,加快草业科技成果的推广与转化。 相似文献