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1.
牛传染性鼻气管炎病毒TK基因缺失株的构建 总被引：8，自引：1，他引：7
将含有牛传染性鼻气管炎病毒（ Ｉ Ｂ Ｒ Ｖ） Ｔ Ｋ 基因的片段克隆于ｐ Ｂｌｕｅｓｃｒｉｐｔ Ｓ Ｋ 中, 再用 Ｂｇ１ Ⅱ和 Ｓａｃ Ｉ切去 Ｔ Ｋ 基因中３４７ｂｐ 的片段, 获得了含缺失 Ｔ Ｋ 基因的重组质粒, 然后插入 Ｌａｃ Ｚ 报告基因, 构建成转移载体。以脂质体转染法将此转移载体与 Ｉ Ｂ Ｒ Ｖ Ｂａｒｔｈａ 株在牛肾细胞（ Ｍ Ｄ Ｂ Ｋ） 上共转染, 经过蓝色蚀斑筛选和蚀斑克隆纯化, 获得了能稳定遗传的重组 Ｉ Ｂ Ｒ Ｖ。经 Ｐ Ｃ Ｒ 和 Ｓｏｕｔｈｅｒｎ 印迹杂交鉴定, 证实该重组病毒为 Ｔ Ｋ 基因缺失突变株。 相似文献
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鸡痘病毒282E4弱毒株TK基因限制性酶切图谱分析 总被引：2，自引：1，他引：1
以限制性内切酶ＢａｍＨＩ、Ｘｂａｌ、Ｃｌａｌ、Ｈ１ｎｄⅢ、Ｎｃａｌ对含有鸡痘病毒（ＦＰＶ）２８２Ｅ４弱毒株３．７ｋｂＨｉｎｄⅢＴＫ基因片段的重组质粒ｐＳＬ１进行单酶和它们之间双酶酶切。结果表明：３．７ｋｂＨｉｎｄⅢＴＫ基因片段上有２个Ｃｌａｌ切点，２个Ｘｂａｌ切点，１个Ｎｃａｌ切点，没有ＢａｍＨＩ切点。随后，用澳大利亚ＦＰＶ２．２ｋｂＨｉｎｄⅢ＋ＣｌａｌＴＫ基因作探针，对各种酶切片段进行Ｓｏｕｔｈｅｒｎ印迹杂交，进一步确定ＴＫ基因位于２．２ｋｂＨｉｎｄⅢ＋Ｃｌａｌ片段中。杂交结果与限制性酶切图谱分析结果相一致。该基因的限制性酶切图谱与澳大利亚ＦＰＶ疫苗株的基本相同。 相似文献
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从包含伪狂犬病病毒（ＰＲＶ）闽Ａ株ＢａｍＨＩ－７片段的重组质粒ｐＰＲ１２８中分离出含有完整糖蛋白ｇｐ５０基因的２．１ｋｂＤＮＡ片段，用ＫｐｎＩ和ＳｔｕＩ酶切后，将其酶切片段分别克隆到ｐＵＣ１９载体中，构建了２．１ｋｂ片段完整测序用质粒。对其序列进行分析，发现与文献报道结果一致，证明分离的ｇｐ５０基因是正确的。将包含ｇｐ５０基因的２．１ｋｂ和１．６ｋｂＤＮＡ片段分别插入带有痘苗病毒天坛株ＴＫ基因区段的ｐＧＪＰ－５质粒Ｐ７．５启动子的下游，构建了ｐＧＢＴ５０－３６和ｐＧＢＴ５０－Ｓ２２个嵌合载体。将嵌合载体通过磷酸钙共沉淀法转染预先感染ＴＫ＋痘苗病毒天坛株的人ＴＫ－１４３细胞或ＣＶ－１细胞，进行体内同源重组。经蚀斑纯化，在ＢｄＵＲ选择压力下，通过光敏生物素标记的探针杂交，获得带有ＰＲＶｇｐ５０基因的重组痘苗病毒。用ＥＬＩＳＡ检测，重组痘苗病毒有特异性ＰＲＶｇｐ５０抗原存在。 相似文献
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猪繁殖和呼吸综合征病毒基质膜蛋白和核衣壳蛋白基因的克隆与鉴定 总被引：3，自引：1，他引：2
猪繁殖和呼吸综合征病毒（ＰＲＲＳＶ）的基质膜（Ｍ）蛋白和核衣壳（Ｎ）蛋白是２种重要的结构蛋白。本试验根据ＰＲＲＳＶ美洲毒株的基因序列,设计了能够扩增Ｍ和Ｎ蛋白基因的１对引物,并在２个引物的两端分别加入ＥｃｏＲＩ和ＢａｍＨＩ酶切位点,经ＲＴ－ＰＣＲ技术获得了一约９５０ｂｐ的目的基因片段,并将此基因克隆于ＰＢＶ２２０载体,构建了ＭＮ蛋白基因重组质粒ＰＢＶＭＮ,进一步由重组质粒回收ＭＮ基因片段亚克隆于ｐＳＫ＋（ｐＢｌｕｅｓｃｒｉｐｔＳＫ＋）测序载体,构建了重组质粒ＰＢＳＭＮ,经部分序列分析鉴定,重组质粒含ＭＮ蛋白基因。此基因的克隆为进一步研究该病毒ＭＮ蛋白免疫学特性及其分子基础和基因诊断技术奠定了基础。 相似文献
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鸡痘病毒282E4株2.2kbTK基因序列分析 总被引：7，自引：2，他引：5
将鸡痘病毒２８２Ｅ４弱毒株基因组中的３．７ｋｂＨｉｎｄⅢ片段克隆到ＫＳ（－）质粒中，亚克隆后，获得含ＴＫ基因正反方向插入的２．２ｋｂＨｉｎｄⅢ－ＣｌａＩ片段的质粒ｐＫＦＡ和ｐＫＦＢ，进而构建出正反方向的系列嵌套缺失质粒。用Ｔ７、Ｔ３引物所做的核苷酸序列分析表明，２．２ｋｂＨｉｎｄⅢ－ＣｌａＩ ＴＫ基因序列长为２１５９ｂｐ，含有两个完整的读码框架（ＯＲＥ）Ｘ和ＴＫ，并确证了ＴＫ基因内存在单一酶切点Ｎ 相似文献
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狂犬病病毒糖蛋白基因在原核细胞中的表达 总被引：2，自引：0，他引：2
将狂犬病病毒糖蛋白（ＲＶｇｐ）基因ＢｇｌⅡ片段（１６７５ｂｐ）分别正向插入到原核高效表达载体ｐＥＴ－１７ｂ和ｐＥＴ－１７ｂ２（用ＳａｃⅠ－ＮｄｅⅠ缺失掉ｐＥＴ－１７ｂ６０ｂｐ含起始密码子ＡＴＧ小片段）的ＢａｍＨⅠ切点，构建重组质粒ｐＥＴ－１７ｂＲＶｇｐ和ｐＥＴ－１７ｂ２ＲＶｇｐ。将其分别转化表达受体菌Ｅ．ｃｏｌｉＢＬ２１（ＤＥ３）和Ｅ，ｃｏｌｉＢＬ２１（ＤＥ３）ｐｌｙｓＳ．ＩＰＴＧ诱导表达，菌体经超声波裂解处理后ＳＤＳ－ｐＡＧＥ，染色，在分子量约６００００处可见重组质粒表达的较宽的蛋白带，以抗ＲＶｇｐＭｃＡｂ进行Ｗｅｓｔｅｒｎ－ｂｌｏｔ检测，表明该表达蛋白为ＲＶｇｐ。通过扫描显示，表达的ＲＶｇｐ占菌体总蛋白的１０％～１４％，其中ｐＥＴ－１７ｂ２ＲＶｇｐ在Ｅ。ｃｏｌｉＢＬ２１（ＤＥ３）中的表达量最高。 相似文献
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NDV融合蛋白裂解位点基因的重组昆虫杆状病毒表达载体的构建… 总被引：1，自引：0，他引：1
将新城疫病毒（ＮＤＶ）Ｆ４６Ｅ９株和ＬａＳｏｔａＥ４株的融合蛋白裂解位点（Ｆｃ）基因从本实验室构建的重组质粒ｐＵＣＦ４６Ｆｃ和ｐＵＣＬａＦｃ中用ＥｃｏＲＩ和ＳａｌＩ切出。将这２个毒株的Ｆｃ基因定向克隆入昆杆状病毒表达载体ｐＳＸＩＶＶＩＸ３的ＥｃｏＲＩ和ＳａｌＩ位点之间，获得２个毒株的Ｆｃ基因克隆ｐＸ３Ｆ４６Ｆｃ和ｐＸ３ＬａＦｃ。重组质粒用ＥｃｏＲＩ／ＳａｌＩ，ＰｓｔＩ酶切法，ＰＣＲ和核酸探针法进行 相似文献
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以Ｘｂａ Ｉ和Ｅｃｏ ＲＶ双酶消化ｐＢＴ１获得肿瘤坏死因子α（ＴＮＦα）ｃＤＮＡ基因片段，以Ｘｂａ Ｉ和Ｓｍａ Ｉ酶切点定向克隆入质粒载体ｐＣＭＶ４，构建了人ＴＮＦα重组质粒ｐＣＭＶ－ＴＮＦα。采用ＤＮＡ－磷酸钙共沉淀法转染ＣＯＳ７和ＮＩＨ３Ｔ３细胞，收集转染后不同时间的细胞培养上清，检测ＴＮＦα的表达水平和生物学活性。ＥＬＩＳＡ检测表明，两种细胞转染后，其培养上清中均有ＴＮＦα抗原存在，而载体等 相似文献
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Marco I Ferroglio E López-Olvera JR Montané J Lavín S 《Veterinary parasitology》2008,152(3-4):321-324
Antibodies to Neospora caninum were determined in the red fox (Vulpes vulpes) in the Pyrenees, north-eastern Spain. Sera from 53 red foxes (29 male and 24 female) were tested using a Neospora agglutination test (NAT). Seroprevalence at dilutions of 1/40, 1/80 and 1/160 was 69.8%, 47.2% and 7.5%, respectively. Sex differences were significant only at a dilution of 1:40. The high seroprevalence observed in red fox suggests that this species is highly exposed to N. caninum in this area. 相似文献
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Hove, K. and H. S. Hansen: Reduction of radiocesium transfer to animal products using sustained release boli with ammoniumiron (III) - hexacyanoferrate (II). Acta vet. scand. 1993,34,287-297.– A sustained release bolus with the cesium binder ammoniu-miron(III) - hexacyanoferrate (AFCF) has been developed as a countermeasure for small ruminants grazing pastures contaminated by radiocesium (134Cs+137Cs). The boli (40-50 g) are produced by compression of a mixture of AFCF, barite and wax. The release of AFCF from boli labelled with 137Cs-iron-hexacyanoferrate complex was studied in laboratory sheep. The release rate followed first order kinetics during the 108 d of observation and decreased from 40 to 22, 110 to 35 and 280 to 25 mg d−1 in sheep treated with 1, 2 or 3 boli respectively. The efficiency of boli in reducing radiocesium transfer to meat and milk was tested in laboratory studies with goats fed 134Cs tracer. Until 40 d after treatment the transfer of radiocesium to milk was reduced by 35%, 60% and 85% in goats given 1, 2 or 3 boli, respectively. The reduction in radiocaesium transfer persisted for 90 d but with a lower efficiency. A similar relationship was found between number of boli and the reduction in radiocesium transfer to meat with an observed maximal reduction of 60%. 相似文献
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鸽禽I型副粘病毒油佐剂灭活苗对肉种鸽免疫效果观察 总被引：4，自引：0，他引：4
用鸽Ａ／ＰＭＶ－１ＳＸ株（简称ＳＸ株）制成的油佐剂灭活苗与ＮＤＶ油佐剂灭活苗分别免疫肉种鸽,免疫后２０天抗体水平达到峰值,抗体水平在３ｌｏｇ２以上维持３０天以上。免疫后４０天用鸽ＳＸ株和新城疫强毒对两种疫苗免疫鸽分别进行攻击,两种疫苗免疫组对ＮＤＶ强毒攻击的保护率均为１００％,鸽Ａ／ＰＭＶ－１油佐剂灭活苗免疫组对鸽ＳＸ株攻击的保护率为１００％,而ＮＤＶ油佐剂灭活苗免疫组对鸽ＳＸ株攻击的保护率仅为６６７％。鸽Ａ／ＰＭＶ－１油佐剂灭活苗田间试验抽检４８份肉种鸽血清,抗体平均值为４３０±１１６ｌｏｇ２。 相似文献
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Javier Del Angel-Caraza Orlando Chávez-Moreno Sergio García-Navarro Carlos Pérez-García 《Journal of veterinary diagnostic investigation》2008,20(5):682-683
Urolithiasis has been reported in veterinary literature in some species of the Mustelidae family, including ferrets. In this species, the most common form of urolithiasis is that caused by struvite. The current study examined the case of mixed urolith in an 8-month-old female ferret (Mustela putorius furo) with antecedents of strangury and hematuria. A cystotomy was carried out to remove the urolith, which had a rough surface and a homogeneously porous interior and was formed by a mixture of struvite (60%) and calcium oxalate dehydrated (40%). 相似文献
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Forty-seven nesting green turtles (Chelonia mydas) were examined for parasitic pathogens. Four species of cardiovascular flukes (Digenea: Spirorchiidae), Learedius learedi, Hapalotrema postorchis, Monticellius indicum and Amphiorchis solus were collected from 39 of 40 of the turtles, and Neospirorchis species were identified in seven of the 47 by histological examination. The pathological changes associated with the spirorchiids and their eggs included aneurysms, arteritis of great vessels with dark nodular thickenings, endocarditis, haemorrhagic lesions, thrombi, vasculitis, and granulomatous reactions in all the turtles. Ozobranchus branchiatus (Hirudinea: Ozobranchidae) leeches were found on the skin of 27 of the 47 turtles and were associated with traumatic purulent and ulcerative dermatitis. Oesophagitis associated with Rameshwarotrema uterocrescens (Digenea: Pronocephalidae) was recorded in 11 of 43 of the turtles. Petechial haemorrhages and unspecific crater-like ulcerated lesions on the gastric mucosa associated with Charaxicephaloides species and Charaxicephalus robustus (Digenea: Pronocephalidae) were observed in four of 40 of the turtles. Cholycystitis and ectasia of mucosal glands associated with Rhytidodoides similis (Digenea: Rhytidodidae) were observed in one of the 47 turtles. Fibropapillomas were observed on the skin of one turtle and fibromas on the skin of two of them. 相似文献
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Pratelli A Cavalli A Normanno G De Palma MG Pastorelli G Martella V Buonavoglia C 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2000,47(4):273-276
The results of vaccination trials carried out on pups with maternally derived antibodies (MDA) to canine parvovirus (CPV), using a modified-live CPV-2b variant vaccine (29-97/40 strain), are reported. The vaccine was able to overcome the obstacle of MDA, and to elicit protective immunity in 100% of the pups whose antibody titres were 1:10-1:40, 83% of the pups with titres of 1:80, 57% of the pups with titres of 1:160, and even in 60% of the pups with antibody titres of 1:320. 相似文献
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Marcos Cláudio Pinheiro Rogério Eliane Minervina de Castro Espedito Cezário Martins Jomar Patrício Monteiro Kleibe de Moraes Silva Magno José Duarte Cândido Tereza Cristina Lacerda Gomes Antoine Francis Roux Bloc Ângela Maria de Vasconcelos Eneas Reis Leite Hélio Henrique Araújo Costa 《Tropical animal health and production》2012,45(1):259-266
This study compares both versions of the nutritional requirement system determined by the National Research Council (NRC) version 1985 (NRC85) and NRC version 2007 (NRC07), for finishing lambs in feedlots. Nineteen crossbred lambs were divided in four groups representing four experimental treatments: one diet according to NRC85 and three diets according to NRC07. The diets recommended by NRC07 considers crude protein intake relative to ruminal undegradable protein at 20, 40, and 60 % levels (NRC07/20, NRC07/40, and NRC07/60). Diets were composed of Brazilian semi-arid native grass silage, soybean meal, corn, annatto byproduct, and limestone. Purchases and sales of lambs were done according to average market prices in Brazil. The economic indicators considered pointed that all treatments were viable but NRC07/20 and NRC07/60 were more profitable with similar net present values (NPVs) and internal return rates (IRRs). NRC07/20 was the best option showing an IRR of 17.20 % and a payback period (PP) of 5.07 considering a fixed annual interest rate of 6 %. Sensitivity analysis considering a 10 % raise in variable costs showed negative NPVs, IRRs inferior to the opportunity cost rates adopted and PPs that exceeded the planning horizon of 7 years for both NRC85 and NRC07/40. 相似文献
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Mortality among Danish dairy cows was examined using data from the Danish Cattle Database (DCD) and a questionnaire survey. Mortality risk has increased from approximately 2% in 1990 to approximately 3.5% in 1999. The increased mortality was seen for all dairy breeds and all age groups. Mortality among older dairy cows (parity 3 and older) was approximately twice the mortality among younger cows. 30-40% of deaths were during the first 30 days of the lactation. Approximately, 58% of dead dairy cows had been euthanised. Replies from the questionnaire indicate that the proportion of euthanised cows has increased in the past 5 years. In 86% of all deaths (questionnaire survey) a primary reason could be identified; 25% were for locomotor disorders. 相似文献
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In southwestern New Jersey during 1986 and 1987, common barn-owls and pigeons were captured on farmsteads and tested for Toxoplasma gondii antibodies by a modified direct agglutination test. In 1986, 3/28 (10.7%) adult and 0/124 nestling owls tested positive at titers of greater than or equal to 1:40. Additionally, 2/34 (5.9%) pigeons tested had T. gondii antibodies at titer of 1:320. In 1987, 9/38 (27.3%) adult and 18/80 (22.5%) nestling owls tested positive at titers of greater than or equal to 1:25; this includes 3/38 (7.9%) adult and 1/80 (1.3%) nestling owls that tested positive at a titer of 1:50. 相似文献