共查询到20条相似文献，搜索用时 625 毫秒
1.
口蹄疫病毒WFL株基因组全长cDNA的克隆及序列分析 总被引：1，自引：0，他引：1
根据口蹄疫病毒基因组的结构特点以及GenBank上公布的全序列，用DNAMAN分别设计了涵盖整个基因组序列的3对引物，从接种口蹄疫病毒WFL株的细胞培养液中提取了病毒基因组RNA，采用RT-PCR方法和RACE法分别扩增了3条基因片段，并将扩增片段分别与T载体连接，在体外分别进行了5-半分子和3’半分子的构建。最后将5’半分子和3’半分子连接成基因组全长cDNA分子。经PCR鉴定、酶切鉴定及全长cDNA测序，证实成功构建了口蹄疫病毒wFL株基因组全长eDNA分子。序列分析结果表明，供试口蹄疫病毒基因组全长为8155nt，5＇UTR长1059nt；具有一个大的读码框，其核苷酸长度为6969nt。包括201aa的前导蛋白基因和2122aa的聚合蛋白基因；3＇UTR长127nt，包括34nt的polv（A）。 相似文献
2.
3.
利用PCR技术，扩增出口蹄疫病毒（FMDV）VP2基因，并克隆到杆状病毒转移载体pBlueBacHis2A上。用重组质粒pB-VP2与重组病毒同时转染sf9昆虫细胞，获得了重组病毒。经过蚀斑筛选纯化后，感染sf9细胞，表达VP2融合蛋白，分子质量为33ku左右。以牛抗O型FMDV血清为第一抗体，通过Western-blotting和Dot-ELISA鉴定，说明VP2基因在真核表达系统中获得正确表达，且可以与牛抗。型FMDV血清发生特异性反应。 相似文献
4.
从亚洲Ⅰ型口蹄疫病毒中提取总RNA，采用反转录聚合酶链反应获得了长度约为700bp的核苷酸片段，大小与预期长度相符。扩增产物经克隆测序，证明其为VP1基因。将该基因亚克隆到原核表达载体pGEX-4T-1，成功构建了重组表达质粒pGEX-VP1；利用1PTG诱导，表达出了53ku的目的蛋白条带。Western-blotting分析表明，表达产物能与亚洲Ⅰ型口蹄疫病毒抗血清特异性结合。亚洲Ⅰ型口蹄疫病毒结构蛋白VP1在大肠埃希氏菌中表达成功。 相似文献
5.
口蹄疫病毒P1基因编码的结构蛋白是构成口蹄疫病毒衣壳的基础，对诱导动物机体产生中和抗体和其他保护性免疫反应有重要作用。文章综述了口蹄疫病毒结构蛋白的结构、抗原特性以及利用重组P1基因制备口蹄疫新型疫苗和诊断检测制剂的最新研究进展。 相似文献
6.
提取2株A型口蹄疫病毒FMDV-L1和FMDV-L2的RNA，用1对通用引物经RT-PCR扩增出2株病毒VP1基因的DNA片段，将扩增的VP1编码序列克隆到质粒载体pGEM-T Easy中，转入大肠埃希氏菌JM109，得到大量携带目的基因的质粒；经过重组质粒的鉴定、测序获得其核苷酸序列；利用序列分析软件及系统发生树绘制软件对FMDV—L1和FMDV-L2以及作为参考毒株的A22／India／17／77进行序列分析。结果表明，核酸序列中的变异多发区要多于氨基酸序列，氨基酸序列最明显的变异发生在构成FMDV抗原位点1的βG-βH环内，其中毒株FMDV-L1和FMDV-L2 RGD序列中的精氨酸(R)发生了变异，分别变成了亮氨酸(L)和谷氨酰胺(Q)。 相似文献
7.
口蹄疫病毒RNA复制酶基因3D的克隆、表达及纯化 总被引：1，自引：0，他引：1
利用RT-PCR扩增了口蹄疫病毒(FMDV)的RNA复制酶基因，并将其克隆到原核表达载体pET-32a( )中。重组质粒在大肠杆菌中表达后的目的蛋白为可溶性形式，纯化产物用感染A型FMDV的豚鼠康复血清进行Western-blotting检测，结果表明，3D基因得到了正确的表达。 相似文献
8.
A型塞内卡病毒(SVA)是近年来新流行的一种传染性病原，与口蹄疫病毒(FMDV)具有相似的基因组结构，所致临床症状也十分相似。为了研发一种新型的基因工程活载体疫苗，用于预防塞内卡病毒感染和口蹄疫，以SVA全长感染性克隆pSVA-GX01为基础，在SVA 2A与2B基因之间插入O型FMDV的VP1基因，经双酶切及测序鉴定，成功获得了重组质粒pSVA-FMDV-VP1-O。将重组质粒转染BHK-21细胞，拯救获得重组病毒rSVA-FMDV-VP1-O,间接免疫荧光试验鉴定显示，该重组病毒可在细胞中表达O型FMDV的VP1蛋白。对该重组病毒进行体外生长增殖特性以及遗传稳定性分析表明，重组毒株和亲本毒株在BHK-21细胞上具有相似的增殖特性，插入的FMDV VP1基因可在细胞传代过程中稳定存在6代。研究结果为开发以塞内卡病毒为载体构建表达口蹄疫病毒VP1蛋白的重组基因工程活载体疫苗提供了参考。 相似文献
9.
参考GenBank中各个血清型口蹄疫病毒3D、vp1、2A基因的标准序列，设计引物P1／P2和S1／S2。建立用于检测口蹄疫病毒及其利用引物S1／S2克隆片段同源性比较而确定血清型的RT-PCR方法。通过敏感性试验检测，2对引物均可以检测到10TCID50的病毒量；特异性试验的检测，2对引物对正常细胞、牛黏膜病病毒、猪瘟病毒、水疱性口炎病毒、牛传染性鼻气管炎病毒的检测结果均为阴性。利用该方法对病牛的流涎液体、水疱液体、舌皮组织、感染犊牛心脏等组织进行检测初步结果显示：该方法可以对O型和AsiaⅠ型口蹄疫病毒进行特异性检测，能够用于口蹄疫急性及亚临床感染的诊断及流行病学调查。 相似文献
10.
根据Genbank中的O型口蹄疫病毒全基因序列设计了一对扩增口蹄疫病毒VP1基因的引物，用该特异性表达引物从口蹄疫阳性质粒pMD—P1中扩增得到目的基因VP1（639bp）。用相同的限制性内切酶酶切目的基因和表达载体PET32a后构建重组表达载体．转化宿主菌BL21（DE3），经酶切及PCR鉴定筛选出阳性克隆．测序证明目的基因正确插入了表达载体，用不同浓度的IPTG诱导VP1基因的表达，收集菌液进行SDS—PAGE电泳，Westerll—blotting分析蛋白免疫原性。结果表明，VP1结构蛋白在大肠杆菌中表达量较高，表达产物的分子量约为41Ku，并能被口蹄疫阳性血清所识别，经分析表达蛋白约占菌体蛋白的34％。口蹄疫病毒VP1蛋白在大肠杆菌中高效表达且表达产物具有免疫原性。 相似文献
11.
Siqueira AK Ribeiro MG Leite Dda S Tiba MR Moura Cd Lopes MD Prestes NC Salerno T Silva AV 《Research in veterinary science》2009,86(2):206-210
The aim of this study was to compare the prevalence of virulence genes in 158 Escherichia coli strains isolated from 51 clinical cases of UTIs, 52 of pyometra and from 55 fecal samples from healthy dogs by PCR. papC was found in 12 (23.5%) strains isolated from UTIs, 19 (36.5%) from pyometra and 10 (18.2%) from feces. papGII was observed in 3 (5.8%) strains from pyometra, and papGIII in 10 (19.6%) from UTIs, 15 (28.8%) from pyometra and 9 (16.4%) from feces. sfaS was detected in 22 (43.1%) strains from UTIs, 24 (46.1%) from pyometra and 19 (34.5%) from feces. hlyA was observed in 17 (33.3%) strains from UTIs, 18 (34.6%) from pyometra and 7 (12.7%) from feces, while cnf-1 was detected in 11 (21.6%) from UTIs, 21 (40.4%) from pyometra and 9 (16.4%) from feces. iucD was observed in 12 (23.5%) strains from UTIs, 9 (17.3%) from pyometra and 1 (1.8%) from feces. usp was found 17 (33.3%) isolates from UTIs and 36 (69.9%) from pyometra. 相似文献
12.
The helminth species composition and helminth burdens of 4 grey duikers, 12 bushbuck, 2 nyala, 2 giraffe, a steenbok, an oribi, a waterbuck and a tsessebe from the Kruger National Park (KNP); of a steenbok and a greater kudu from the farm Riekerts Laager, Transvaal; of a single blue duiker from the Tsitsikama Forest National Park, and of a blue wildebeest, a red hartebeest, a gemsbok and 2 springbok from the Kalahari Gemsbok National Park (KGNP) were collected, counted and identified. New parasite records are: Agriostomum equidentatum from the gemsbok, Cooperia neitzi from the bushbuck, Cooperia sp. from the gemsbok and the red hartebeest, Cooperia yoshidai from the waterbuck and the tsessebe, Dictyocaulus viviparus from the bushbuck, Haemonchus bedfordi from the waterbuck, Haemonchus contortus from the gemsbok, Haemonchus krugeri from the steenbok from the KNP, Impalaia nudicollis from the gemsbok and the red hartebeest, Impalaia tuberculata from the oribi and the waterbuck, Impalaia spp. from the kudu, Longistrongylus meyeri from the steenbok from Riekerts Laager and the gemsbok, Longistrongylus sabie from the steenbok from the KNP, Longistrongylus schrenki from the tsessebe, Parabronema sp. from the tsessebe and the red hartebeest, Paracooperia serrata from the gemsbok and the steenbok from the KGNP, Pneumostrongylus calcaratus from the bushbuck, Strongyloides sp. from the gemsbok, Trichostrongylus sp. from the gemsbok, the red hartebeest and the steenbok from the KGNP, Trichostrongylus axei from the blue duiker, Trichostrongylus falculatus from the bushbuck and the oribi, Trichostrongylus instabilis from the bushbuck, the steenbok from the KNP and the oribi and Trichostrongylus thomasi from the grey duikers and tsessebe. Host specificity of the parasites was not marked and crossinfestation was common. This was not true for the giraffe, since none of the helminths of these animals were found in the antelope and vice versa. 相似文献
13.
Hallstein Grnstl 《Acta veterinaria Scandinavica》1980,21(1):11-17
The udders from 13 culled ewes and liver, spleen, kidney, lung and brain from 15 lambs, 11 months old, were examined for the presence of Listeria monocytogenes (Lm) at slaughter. Lm was isolated from 1 of 13 udders, from 6 of the 15 brains and from 0–4 of the other organs from each of the 15 lambs.Internal organs from 68 sheep submitted for post-mortem examination were examined in the same way. Lm was isolated from 25 of these animals. Lm was isolated from the brain of 7 of 9 animals with encephalitis, and from 0–3 of the other 4 organs examined. Lm was also isolated from 10–20 % of the organs from animals with other diagnoses. Altogether 9 of 10 animals with encephalitis and 16 of 58 with other diagnoses (28 %) were found to harbour this organism. 相似文献
14.
欧氏和玛氏对盲囊线虫线粒体nad1基因部分序列的比较研究 总被引：1，自引：0，他引：1
欧氏对盲囊线虫(Contracaecum ogmorhini)和玛氏对盲囊线虫(Contracaecum margolisi)是属于欧氏对盲囊线虫复合种中的两个种，前来自于南半球，后来自于北半球，二在地理分布和宿主特异性方面均有明显的不同。本研究用γ^33P对引物进行标记，通过聚合酶链式反应一单链构象多态性(PCR—SSCP)分析和DNA序列分析技术对来自南半球两个不同地理种群的欧氏对盲囊线虫和北半球种群的玛氏对盲囊线虫在线粒体NADH脱氢酶亚单位Ⅰ基因(nad1)部分序列的差异及种群遗传关系进行了研究。结果显示：南半球两个种群的序列相似性很高，而北半球种群与南半球种群的序列相似性则相对较低，它们之间had1部分序列种间遗传差异大于种内，且存在着2个可作为区分两的遗传标记。种群遗传关系分析也表明：北半球的玛氏对盲囊线虫种群作为一个独立的新种而区别于南半球的两个欧氏对盲囊线虫种群。 相似文献
15.
AIMS: To use multilocus enzyme electrophoresis to determine the genetic structure of Staphylococcus intermedius from normal skin of dogs and those isolated from a variety of disease conditions and to distinguish clinically important strains in dogs. METHODOLOGY: The diversity amongst 129 isolates of S intermedius from the skin and mucosa of 32 healthy dogs and 120 isolates from diseased sites in 120 individual dogs was examined using multilocus enzyme electrophoresis. Associations among ETs were examined to determine the diversity of isolates. RESULTS: Twenty two ETs were distinguished comprising 21 containing isolates from diseased sites and 11 containing isolates from normal dogs. The majority of isolates (171 of 249; 69% were located in two ETs (ET1 and ET 4), that were not distinguishable phenotypically. ET 1 contained 94 isolates (54 isolates from healthy dogs and 40 isolates from diseased sites) and ET 4 contained 77 isolates (46 from healthy dogs and 31 isolates from diseased sites). Further, 77.5% of isolates from healthy dogs were present in ET 1 and ET 4 and 59% of isolates from diseased dogs belonged to the same two ETs. There was only a small difference in genetic diversity among isolates taken from healthy dogs (11 ETs; H = 0.182) and those isolates taken from clinical specimens from diseased dogs (21 ETs; H = 0.218). Of the 21 ETs from diseased sites, ET 16 contained all six isolates from Staphylococcal Scalded Skin Syndrome in racing Greyhounds. CONCLUSIONS: The small difference in genetic diversity between isolates from the skin and mucosa of healthy dogs and isolates from various diseases, as well as the presence of the majority of isolates in two ETs, is consistent with the role of S intermedius as an opportunistic pathogen. The confinement of all Staphylococcal Scalded Skin Syndrome isolates within one ET is confirmation of this entity as a distinct disease of dogs. 相似文献
16.
Goldman CG Matteo MJ Loureiro JD Degrossi J Teves S Heredia SR Alvarez K González AB Catalano M Boccio J Cremaschi G Solnick JV Zubillaga MB 《Veterinary microbiology》2009,133(3):287-291
The mechanism by which Helicobacter species are transmitted remains unclear. To examine the possible role of environmental transmission in marine mammals, we sought the presence of Helicobacter spp. and non-Helicobacter bacteria within the order Campylobacterales in water from the aquatic environment of marine mammals, and in fish otoliths regurgitated by dolphins. Water was collected from six pools, two inhabited by dolphins and four inhabited by seals. Regurgitated otoliths were collected from the bottom of dolphins' pools. Samples were evaluated by culture, PCR and DNA sequence analysis. Sequences from dolphins' water and from regurgitated otoliths clustered with 99.8-100% homology with sequences from gastric fluids, dental plaque and saliva from dolphins living in those pools, and with 99.5% homology with H. cetorum. Sequences from seals' water clustered with 99.5% homology with a sequence amplified from a Northern sea lion (AY203900). Control PCR on source water for the pools and from otoliths dissected from feeder fish were negative. The findings of Helicobacter spp. DNA in the aquatic environment suggests that contaminated water from regurgitated fish otoliths and perhaps other tissues may play a role in Helicobacter transmission among marine mammals. 相似文献
17.
18.
Aoi SUMIYOSHI Koichi KITAO Takayuki MIYAZAWA 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2022,84(1):157
Foamy viruses have been isolated from various mammals and show long-term co-speciation with their hosts. However, the frequent inter-species transmission of feline foamy viruses (FFVs) from domestic cats to wild cats across genera has been reported. Because infectious molecular clones of FFVs derived from wild cats have not been available, whether there are specific characteristics enabling FFVs to adapt to the new host species is still unknown. Here, we obtained the complete genome sequences of two FFV isolates (strains NV138 and SV201) from leopard cats (Prionailurus bengalensis) in Vietnam and constructed an infectious molecular clone, named pLC960, from strain NV138. The growth kinetics of the virus derived from pLC960 were comparable to those of other FFVs derived from domestic cats. Phylogenetic analysis revealed that these two FFVs from leopard cats are clustered in the same clade as FFVs from domestic cats in Vietnam. Comparisons of the amino acid sequences of Env and Bet proteins showed more than 97% identity among samples and no specific amino acid substitutions between FFVs from domestic cats and ones from leopard cats. These results indicate the absence of genetic constraint of FFVs for interspecies transmission from domestic cats to leopard cats. 相似文献
19.
光生物素标记的口蹄疫病毒cDNA探针的制备及其对口蹄疫病毒RNA检测的试验研究 总被引：1，自引：0，他引：1
本文首次报道了利用pF1034质粒制备FMDV O型特异性探针,并通过PCR反应扩增FMDO_1K株病毒基因组的第2962位与3071位之间共110bp序列,制备了能检测O型、A型和亚洲I型FMDV RNA的群(组)特异性探针。用硝酸纤维素膜斑点杂交试验表明,二者均能检测出10pg水平的O_9K毒株的纯RNA;但前者只与O型FMDV RNA杂交,与A型及亚洲I型FMDV RNA无交叉杂交现象;而后者则能与O型、A型和亚洲I型的FMDV RNA发生杂交反应。对照试验显示:此两种探针与SVDV ssRNA、BTV dsRNA、EHDV dsRNA、DHV ssRNA、PRV DNA、乳鼠组织细胞RNA、BHK_(21)克隆13细胞RNA及DNA等均不出现交叉杂交现象,但与IBR DNA有假阳性杂交反应。 相似文献