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用PCR方法,成功扩增出国内分离株禽流感病毒H9N2型非结构蛋白(NS1)基因cDNA,并将该片段连接到pMD 18-T载体上,转化TGI感受态细胞,在含氨苄青霉素的LB平板上筛选阳性克隆,提取质粒,经限制性内切酶鉴定,对目的片段进行测序,结果获得了NS1基因全长,约700bp.该片段的核酸序列与已知A/Chick/HongKong/1074/99(H9N2)、A/Chick/HongKong/1073/99(H9N2)毒株序列进行同源性比较,同源性皆为94.37%,氨基酸序列同源性分别为93.91%和93.48%. 相似文献
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为了研究H6亚型禽流感病毒的分子特征,本研究从鸭的咽喉拭子样本中分离到一株禽流感病毒,通过血凝抑制试验证实该分离株为H6亚型流感病毒,命名为A/duck/Heilongjiang/QG 1/2013(H6).对其血凝素基因进行了克隆和序列分析,结果表明分离株与我国鸭源禽流感病毒A/duck/Guangxi/585/2005 (H6N5)的同源性最高,达到97.8％.系统发育树分析进一步证实分离株与A/duck/Guangxi/585/2005 (H6N5)的亲缘关系最近,共同起源于台湾分离株A/chicken/Taiwan/G23/87 (H6N1).本研究为H6亚型禽流感的流行病学研究补充了新的数据. 相似文献
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一株野鸭源H4N6亚型禽流感病毒A/mallard/Yancheng/2005的全基因组克隆和序列分析 总被引：1，自引：0，他引：1
应用流感病毒通用引物对盐城珍禽自然保护区野鸭泄殖腔棉拭子分离株禽流感病毒A/mallard/Yancheng/2005（简称Mallard/YC/2005）（H4N6）进行全基因组序列扩增,结合GenBank中的相关序列进行遗传进化分析。结果表明,Mallard/YC/2005（H4N6）HA基因与A/duck/Siberia/1701/1996（H4N6）的核苷酸同源性最高（97.5%）,推导的氨基酸剪切位点序列为PEKASR,为典型低致病性禽流感病毒的特征序列;神经氨酸酶（NA）、非结构蛋白（NS）均没有氨基酸缺失;基质蛋白2（M2）与宿主特异性有关的位点及碱性聚合酶2（PB2）的627位都是亲禽类细胞的氨基酸;M基因与家鸭分离株A/duck/Yangzhou/02/2005（H8N4）的核苷酸同源性为99.4%,PA基因与家鸭分离株A/duck/Jiangxi/1286/2005（H5N2）的核苷酸同源性达98.9%,说明这2个基因已经在家鸭体内存在并参与了基因重排。 相似文献
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采用常规血清学试验和特异性RT-PCR方法,对我国华东地区家养水禽流感病毒进行了长达4年的跟踪监测,分离到多株H1亚型禽流感病毒,并对其中的1株A/Duck/Yangzhou/163/2003(简称Dk/YZ/163/03)的血凝素基因进行了序列测定,并与GenBank中收录的其他序列进行了比较。结果发现,Dk/YZ/163/03的血凝素基因(HA)与禽流感日本分离株A/Japan/91(H1)的同源性最高;而神经氨酸酶基因(NA)与Mallard dk/A1b/35/1976(H1N1)同源性最高;其推导的氨基酸剪切位点序列为P-S-I-Q-S-R,是典型低致病性禽流感病毒的特征序列,这与该毒株对SPF鸡和BALB/c小鼠的低致病特性相吻合。 相似文献
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禽流感病毒A/Chicken/Yangzhou/N/2005(H9N2)HA基因的克隆及序列分析 总被引：1，自引：0，他引：1
根据GenBank公开发表的H9N2型禽流感病毒血凝素(HA)基因序列设计1对引物,用RT-PCR方法成功扩增出H9N2型禽流感病毒分离株A/Chicken/Yangzhou/N/2005(H9N2)HA基因,凝胶电泳结果显示,扩增产物为1.7 kb的单一条带,与预期结果相符.将PCR扩增片段连接到pCR2.1-T载体上,重组质粒酶切鉴定正确.序列测定分析结果显示,所获序列与GenBank中收录的H9N2亚型分离株核苷酸同源性最高达99%,与原型毒株A/Furkey/Wisconsin/1/1966(H9N2)氨基酸同源性为89%.对HA裂解位点附近的氨基酸序列分析表明,此毒株为低致病力毒株. 相似文献
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为了解禽流感病毒在华东地区家禽体内的进化情况,作者对华东地区某活禽市场的家禽进行了禽流感病毒的流行病学监测,并分离鉴定出1株H11N2亚型禽流感病毒A/duck/Jiangxi/k0701/2009。对该株病毒全基因组的遗传进化分析表明A/duck/Jiangxi/k0701/2009的8个基因片段来源广泛,与亚洲及欧洲某些地区的毒株存在广泛的重组现象。其PB2基因和NS基因可能由A/mallard/Korea/GH170/2007(H7N7)提供,NA基因与高致病性禽流感病毒A/duck/Hebei/0710/2009(H5N2)的NA基因亲缘关系较近,而PB1、PA和NP基因则可能与A/Muscovy duck/Thailand/CU-LM1984/2009(H4N9)有共同来源。本研究结果提示该株病毒可能为1株多元重组病毒,且可能在高致病性禽流感病毒的进化中起了重要作用。 相似文献
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两株不同亚型禽流感病毒NS1基因的表达及抗原性检测 总被引：3，自引：0，他引：3
为进一步研究禽流感病毒非结构蛋白NS1的功能及其在临床中的应用,本试验分别对H5N1、H9N2两株不同亚型禽流感病毒的NS1基因进行了原核表达及抗原性检测。实验分别用限制性内切酶EcoRI和XhoI消化不同亚型NS1基因的重组质粒pMD-18T-NS1(H9N2)和pMD-18T-NS1(H5N1),获得目的片段NS1,然后将NS1基因克隆到原核表达载体pProEXHTc中。将重组质粒pProc-NS1(H9N2、H5N1)转化DH5α感受态细胞,用1mmol/LIPTG诱导表达,表达产物进行SDS-PAGE检测,目的蛋白分子量大小为30ku,与预期结果相符。West-ern-blot检测表明所表达的蛋白能与NS1的多克隆抗血清反应,并且存在交叉反应。 相似文献
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一株鸡源H6N1亚型禽流感病毒全基因的分子特征 总被引：2，自引：2，他引：0
2008年国家禽流感参考实验室在我国禽流感流行病学调查期间分离到1株鸡源H6N1亚型禽流感病毒(AIV)A/Chicken/ZheJiang/80/2008(H6N1)(简称为CK/ZJ/80/08),为了弄清该病毒的分子特征,我们对其8个基因片段分别进行扩增和序列测定,对每个基因进行BLAST分析,找出同源性最高的毒株。利用DNAStar中的Megalign功能进行进化分析。结果表明CK/ZJ/80/08的HA裂解位点附近的氨基酸序列为QIETR↓GLF,推测可能为一株低致病力AIV。其HA基因与日本北海道的A/duck/Hokkaido/228/2003(H6N8)和黑龙江的A/mallard/Heilongjiang/131/2006(H6N2)以及香港早期分离株A/chicken/HongKong/17/77(H6N1)等处于同一分支;NA基因在颈部没有缺失,与A/duck/Tsukuba/718/2005(H1N1)、A/goose/Guangdong/1/96(H5N1)等处于同一分支;M基因与A/duck/Hokkaido/W90/2007(H10N7)高度同源(同源性为99%);NS基因与A/duck/Denmark/65047/04(H5N2)和A/goose/Guangdong/1/96(H5N1)处于同一分支。NP、PA、PB1、PB2分别与贵州和江西分离的H5N2亚型AIV的相应基因关系密切,同源性分别为98%、97%、97%、97%。由此推测CK/ZJ/80/08可能是由H6N2、H1N1、H10N7、H5N2等多个亚型病毒重组而成。 相似文献
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The duodenum of various ruminants was examined by immunohistochetnical staining for serotonin, substance P and Metenkephalin-Arg6-Gly7-Leu8 (MENK8). Serotonin- and substance P-immunoreactive endocrine cells were detected in samples from cow, calf, sheep, goat, and barbary sheep. MENK8-immunoreactive endocrine cells were detected exclusively in samples from cow and calf. Substance P and MENK8 immunoreactivity was found in serotonin-immunoreactive endocrine cells of cattle. Substance P- and MENK8-immunoreactive nervous elements were detected in all ruminants examined. The present results suggest that the expression and/or the mechanism that controls the expression of each peptide might differ between endocrine cells and nerve cells and might also depend on animal species. 相似文献
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马流感A/马/京防/74-1(H7N7)毒株HA基因的序列分析 总被引：6，自引：0，他引：6
流感病毒(Influenza Virus)根据血凝素(HA)和神经氨酸酶(NA)两种表面抗原蛋白分为不同的亚型,马流感I型(H7N7)和Ⅱ型(H3N8)是其中比较重要的两个亚型.本研究应用无特定病原体(SPF)鸡胚增殖马流感病毒A/马/京防/74-1(H7N7)毒株,TRIzol LS Reagent提取病毒RNA,RT-PCR扩增HA基因全片段,克隆到PMD18-T载体上,并进行了鉴定和序列测定.所获得的HA基因片段长1 727 bp,编码563个氨基酸残基.根据推导的氨基酸序列进行预测,有7个潜在的糖基化位点和16个半胱氨酸残基,通过序列分析推断,A/马/京防/74-1(H7N7)株病毒的HA来源于禽类,是一株通过基因重排出现的重组病毒. 相似文献
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Godfroid J Scholz HC Barbier T Nicolas C Wattiau P Fretin D Whatmore AM Cloeckaert A Blasco JM Moriyon I Saegerman C Muma JB Al Dahouk S Neubauer H Letesson JJ 《Preventive veterinary medicine》2011,102(2):118-131
Following the recent discovery of new Brucella strains from different animal species and from the environment, ten Brucella species are nowadays included in the genus Brucella. Although the intracellular trafficking of Brucella is well described, the strategies developed by Brucella to survive and multiply in phagocytic and non-phagocytic cells, particularly to access nutriments during its intracellular journey, are still largely unknown. Metabolism and virulence of Brucella are now considered to be two sides of the same coin. Mechanisms presiding to the colonization of the pregnant uterus in different animal species are not known. Vaccination is the cornerstone of control programs in livestock and although the S19, RB51 (both in cattle) and Rev 1 (in sheep and goats) vaccines have been successfully used worldwide, they have drawbacks and thus the ideal brucellosis vaccine is still very much awaited. There is no vaccine available for pigs and wildlife. Animal brucellosis control strategies differ in the developed and the developing world. Most emphasis is put on eradication and on risk analysis to avoid the re-introduction of Brucella in the developed world. Information related to the prevalence of brucellosis is still scarce in the developing world and control programs are rarely implemented. Since there is no vaccine available for humans, prevention of human brucellosis relies on its control in the animal reservoir. Brucella is also considered to be an agent to be used in bio- and agroterrorism attacks. At the animal/ecosystem/human interface it is critical to reduce opportunities for Brucella to jump host species as already seen in livestock, wildlife and humans. This task is a challenge for the future in terms of veterinary public health, as for wildlife and ecosystem managers and will need a "One Health" approach to be successful. 相似文献
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根据GenBank中发表的GPMV SF02株的NP基因的核苷酸序列设计合成一对引物,采用RT-PCR扩增出与预期设计的1470bp大小相符的片断,将此扩增产物克隆入PMD18-T载体,进行序列测定和分析.结果表明:所扩增的NP基因的核苷酸长为1 470bp,共编码490个氨基酸.同源性分析表明:JS/1/97/Go与国内的SF02株的NP基因核苷酸同源性为98.6%,氨基酸同源性为99.6%.由此可见,鹅副粘病毒JS/1/97/Go分离株与SF02株的亲缘关系较近,同属于新城疫Ⅶ型基因.而其与LaSota株的核苷酸同源性为85.2%,氨基酸同源性为90.8%.说明该毒株相对于经典的NDV在NP基因上已发生了较大的变异. 相似文献
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J Houttu 《Equine veterinary journal》1991,23(3):163-165
This paper presents the results of arthroscopic surgery in 42 Standardbred trotters and three Finnish horses. Forty-five horses were operated on. The age range was one to seven years; 73 per cent of the horses were three years old or younger. Horses showed a variety of clinical signs ranging from moderate to severe lameness at slow speeds, to obscure lameness manifesting only at high speeds. Synovial effusion of the fetlock joint was rare. In this series, 44 horses that had Type I fragments and one horse that had Type III fragments were operated on. Of the 45 horses operated on, 23 (51 per cent) returned to speed training in three months and 41 (91 per cent) returned to speed training in six months. Three of 45 (6 per cent) were lame three months after the surgery when the trainer attempted to start speed training. One of those was sound six months after the operation. The remaining two (4 per cent) stayed lame due to a lesion in the affected joint. Two of 45 (4 per cent) discontinued training for other reasons. The horse with Type III fragments returned to speed training in three months. 相似文献
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MARTIN HOFSTETTER DVM PHILIPPE GÉDET Diplom-HTL-Ingenieur MARCUS DOHERR Diplomate ECVPH STEPHEN J. FERGUSON PhD FRANCK FORTERRE Diplomate ECVS 《Veterinary surgery : VS》2009,38(1):49-58
Objective— To study the kinematics of cervical spine segment C4 –C5 and its association with disc dimensions and the coupled motion (CM) in relation to primary motion (PM).
Study Design— Cadaveric biomechanical study.
Animals— Cadavers of large breed dogs (>20 kg; n=11).
Methods— Spines were freed from muscles. Radiographs were taken orthogonal to the C4 –C5 disc space and disc thickness, endplate width, and height were measured. Spines were mounted on a simulator for 3-dimensional motion analysis. Data were recorded with an optoelectronic motion analysis system. Range of motion (ROM) and neutral zone (NZ) were determined in the direction of flexion/extension, left/right lateral bending, and left/right axial rotation, as well as the ROM of CM.
Results— ROM in flexion and extension was similar; there was no CM in flexion/extension. Left/right axial rotation and left/right lateral bending were coupled to the same side. CM was 1.72 and 3.56 times the ROM of the PM in lateral bending and axial rotation, respectively. Disc dimensions were positively correlated with body weight. Flexion/extension magnitude was significantly reduced for larger endplates, but axial rotation was not influenced. Lateral bending had no correlation with weight or disc dimensions.
Conclusion— Left/right lateral bending and left/right axial rotation are coupled differently in the C4 –C5 segment in dogs compared with humans.
Clinical Relevance— The canine C4 –C5 spinal segment has unique motion coupling patterns that should be considered for dynamic implant designs. 相似文献
Study Design— Cadaveric biomechanical study.
Animals— Cadavers of large breed dogs (>20 kg; n=11).
Methods— Spines were freed from muscles. Radiographs were taken orthogonal to the C
Results— ROM in flexion and extension was similar; there was no CM in flexion/extension. Left/right axial rotation and left/right lateral bending were coupled to the same side. CM was 1.72 and 3.56 times the ROM of the PM in lateral bending and axial rotation, respectively. Disc dimensions were positively correlated with body weight. Flexion/extension magnitude was significantly reduced for larger endplates, but axial rotation was not influenced. Lateral bending had no correlation with weight or disc dimensions.
Conclusion— Left/right lateral bending and left/right axial rotation are coupled differently in the C
Clinical Relevance— The canine C
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猪流感病毒A/Swine/Inner Mongolia/547/01分离株神经氨酸酶基因的克隆及真核表达 总被引：2，自引：0，他引：2
根据GenBank中登录的猪流感病毒参照序列设计了扩增神经氨酸酶(NA)基因的特异性引物.从猪流感病毒A/Swine/Inner Mongolia/547/01(H3N2)株感染的鸡胚液中直接提取病毒RNA,经RT-PCR扩增后将其克隆到pMD18-T载体上,进行序列测定和拼接.用HindⅢ酶切后,亚克隆到杆状病毒转移载体pMelBacB上,得到重组转移载体pMelBacNA,然后与线性化杆状病毒DNA(Bac-N-BlueTMDNA)共转染于sf9昆虫细胞.经3轮噬斑纯化,提取其DNA经PCR鉴定,获得重组杆状病毒株.SDS-PAGE结果显示,目的基因已获得表达,Western blot和神经氨酸酶活性检测结果显示表达产物具有良好的免疫原性和神经氨酸酶活性,为SIV亚单位疫苗和NA基因的深入研究奠定了基础. 相似文献
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马流感病毒(A/equine/Qinghai/1/94)核蛋白基因的序列测定及同源性分析 总被引：1，自引：0，他引：1
根据已发表的马流感病毒核蛋白基因序列，设计并合成一对特异性引物，经反转录-聚合酶链反应（RT-PCR）成功扩增出了我国马流感病毒（A/equine/Qinghai/1/94核蛋白基因。将该片段连接到PGEM-T-EASY载体并转化DH5α，提取阳性菌落的质粒径EcRo1酶切的PCR鉴定其大小为1.5kb左右，对其测序并进行分析发现，与A/Equine/Kentucky/2/86,A/Equine/Miami/1/63等关系较近，同源率为93.3%--93.4%，而与我国马流感吉林A/Equine/Jilin/1/89株关系较远，同源率仅为84.6%。 相似文献