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1.
为了研究大熊猫轮状病毒VP3基因,为以后研究此基因的生物学功能、建立该病毒的检测方法以及研制新型大熊猫轮状病毒基因疫苗奠定基础,试验采用MA-104细胞培养增殖大熊猫轮状病毒CH-1株,提取总RNA,运用RT-PCR扩增外衣壳蛋白VP3基因,将其与p MD19-T Simple载体连接并转化DH5α,经PCR扩增和测序分析进行鉴定,用生物信息学软件预测其功能,并构建系统进化树。结果表明:成功获得大熊猫轮状病毒VP3蛋白基因,长为2 591 bp,包含1个2 508 bp的开放阅读框,编码835个氨基酸,测序结果在Gen Bank中的登录号为HQ641295。VP3蛋白基因编码产物分子质量为97 937.91 u,最大疏水指数为2.256,最小疏水指数为-3.000;无信号肽序列;无跨膜结构;在线预测VP3有37个抗原表位;预测其可能包含10个N-糖基化位点,1个c AMP-c GMP独立蛋白激酶磷酸化位点,20个蛋白激酶C磷酸化作用位点,18个酪蛋白激酶Ⅱ磷酸化作用位点,1个酪氨酸激酶磷酸化作用位点,5个N-豆蔻酰化位点。大熊猫轮状病毒的VP3基因与猪轮状病毒VP3基因的进化距离最近。  相似文献   

2.
为研究大熊猫轮状病毒(Giant Panda Rotavirus,GPRV)CH-1株外衣壳蛋白VP7基因的结构及功能,用MA-104细胞(恒河猴胎猴肾细胞)增殖GPRV,提取总RNA,运用RT-PCR扩增VP7基因,并测序,用生物信息学软件预测其功能,并构建系统进化树。结果显示成功获得了长1042bp的GPRV VP7蛋白基因(GenBank登录号GU188284)。经生物信息学分析,此序列包含1个981bp的完整开放阅读框,编码326个氨基酸;预测GPRV VP7蛋白理论相对分子质量为37354.8u,等电点为4.76,半衰期为30h,不稳定系数为26.71,总平均亲水性为-0.015,疏水性介于-2.344~3.567;1—24位氨基酸可能是信号肽序列;跨膜结构分析表明VP7蛋白有2个跨膜区,其N端和C端都位于病毒膜外区;抗原表位预测显示VP7蛋白有12个抗原决定簇;结构预测显示其可能包含2个N-糖基化作用位点,5个蛋白激酶C磷酸化作用位点、4个酪蛋白激酶Ⅱ磷酸化作用位点、1个酪氨酸激酶磷酸化作用位点、4个N-豆蔻酰化位点、1个原核膜脂蛋白脂附着点和1个革兰氏阳性球菌表面蛋白‘锚’六肽。系统进化树分析显示GPRV VP7基因与人A组轮状病毒VP7基因的进化距离最近。本研究成功获得了GPRV VP7基因,为今后研究此基因的生物学功能以及建立该病毒的诊断方法奠定基础。  相似文献   

3.
为了了解贵州省猪流行性腹泻(porcine epidemic diarrhea, PED)的流行情况,试验对猪流行性腹泻病毒(Porcine epidemic diarrhea virus, PEDV)贵州流行株(GZ2022)N基因序列进行克隆、测序,并利用DNAStar、ProtParam、ProtScale等生物信息学分析软件对PEDV-GZ2022株N基因的同源性和遗传进化地位,以及N蛋白的分子特征和潜在功能进行分析预测。结果表明:PEDV-GZ2022株N基因大小约为1 364 bp,编码441个氨基酸;PEDV-GZ2022株N基因核苷酸序列与GenBank中的PEDV参考毒株N基因的相似性为95.5%~99.8%,与PEDV经典毒株CV777 N基因核苷酸序列的相似性为95.6%,与SDLY2020毒株N基因核苷酸序列的相似性高达99.8%,PEDV-GZ2022毒株与经典毒株CV777关系相隔较远,与PEDV变异株GⅡ-b位于同一分支;PEDV-GZ2022株N蛋白中含有较多的疏水性氨基酸,为疏水性蛋白;该蛋白质存在4个N糖基化位点和37个O糖基化位点,有31个丝氨酸、...  相似文献   

4.
根据GenBank上发表的副猪嗜血杆菌(HPS)全基因组序列中的外膜蛋白OMP2基因的核苷酸序列,利用Premier Primer5.0和Oligo6.0软件设计1对特异性引物,对从贵州省分离的HPS的OMP2基因进行扩增、克隆、测序和生物信息学分析。测序结果比表明,扩增的HPS分离菌株目的基因长度为1092bp,共编码363个氨基酸,与GenBank上公布的SH0165株(NC011852)中对应的核苷酸序列同源性为99.8%,氨基酸同源性100%。生物信息学分析结果表明,OMP2外膜蛋白的分子质量为39087.45Daltons,理论等电点pI为9.21,不稳定系数为16.73,推测其为稳定性蛋白;脂肪指数为70.63,总体平均亲水性为-0.494,推测该蛋白是一种亲水性蛋白;该蛋白含有α-螺旋、β-折叠、β-转角和无规则卷曲,柔性区域较多,呈散在分布,有8个主要的抗原表位;OMP2外膜蛋白序列最前端的19个N-末端氨基酸残基为信号肽序列,最佳切割位点在19~20个氨基酸,且信号肽区域主要为疏水性氨基酸;该蛋白无跨膜区;预测OMP2外膜蛋白可能含有4个N-糖基化位点、1个cAMP和cGMP依赖性蛋白激酶磷酸化位点、8个蛋白激酶C磷酸化位点、7个酪蛋白激酶II磷酸化位点、1个酪氨酸激酶磷酸化位点、9个N-肉豆蔻酰化位点。  相似文献   

5.
根据GenBank 上公布的副猪嗜血杆菌外膜蛋白OMP5 基因的核苷酸序列,设计1 对特异性引物,对贵州省HPS 分离株OMP5 基因进行扩增。结果表明所扩增的OMP5 基因的长度为1116 bp,与SH0165 株对应的核苷酸序列和氨基酸同源性均为100%。生物信息学分析结果表明,OMP5 基因所编码的外膜蛋白P5 的理论等电点pI为9.34,不稳定系数为20.44,推测其为稳定性蛋白;脂肪指数为83.94,总体平均亲水性为-0.172,推测该蛋白是一种亲水性蛋白;该蛋白含有丰富的α-螺旋、β-折叠、β-转角和无规则卷曲,无跨膜区,最前端的21 个N-末端氨基酸残基为信号肽序列并且主要为疏水性氨基酸,最佳切割位点在第21~22 个氨基酸之间;结构位点预测表明该蛋白含有多个不同的糖基化和磷酸化位点。  相似文献   

6.
根据GenBank上公布的副猪嗜血杆菌外膜蛋白OMP5基因的核苷酸序列,设计1对特异性引物,对贵州省HPS分离株OMP5基因进行扩增。结果表明所扩增的OIvIP5基因的长度为1116bp,与SH0165株对应的核苷酸序列和氨基酸同源性均为lOO%。生物信息学分析结果表明,OMP5基因所编码的外膜蛋白P5的理论等电点pl为9.34不稳定系数为20.44,推测其为稳定性蛋白;脂肪指数为83.94,总体平均亲水性为-0.172,推测该蛋白是一种亲水性蛋白;该蛋白含有丰富的α-螺旋、β-折叠、β-转角和无规则卷曲,无跨膜区,最前端的21个N-末端氨基酸残基为信号肽序列并且主要为疏水性氨基酸,最佳切割位点在第21~22个氨基酸之间;结构位点预测表明该蛋白含有多个不同的糖基化和磷酸化位点。  相似文献   

7.
为了研究Ⅶ型新城疫病毒(Newcastle disease virus,NDV)CY1株的进化情况,分析其遗传变异和基因功能,试验应用RT-PCR扩增NDV辽宁分离株CY1株的融合蛋白(fusion,F)基因,进行测序分析,构建系统进化树,并用生物信息软件对蛋白结构功能进行分析。结果显示,NDV F基因全长1662 bp,编码553个氨基酸。生物信息学分析结果表明,F蛋白的分子质量为58991.0 u,理论等电点为8.48,半衰期为30 h(哺乳动物类网状细胞),不稳定系数31.62,脂肪指数为108.12,总平均疏水性0.174,最大疏水性为3.078,最小疏水性为-2.578,信号肽序列为第1-31位氨基酸残基,跨膜区结构分析结果表明该蛋白为跨膜蛋白,预测可能含有6个N-糖基化作用位点、9个酪蛋白激酶Ⅱ磷酸化位点、7个蛋白激酶C磷酸化位点、14个N-豆蔻酰化位点、1个酰胺化位点、1个亮氨酸拉链、1个微体C端靶信号位点、1个双向核定位信号。系统进化分析结果表明,NDV CY1株与zj1株进化距离最近。  相似文献   

8.
羊源多杀性巴氏杆菌ompW基因的克隆及生物信息学分析   总被引:1,自引:1,他引:0  
试验旨在克隆羊源多杀性巴氏杆菌ompW基因,并对其序列进行生物信息学分析。根据GenBank中多杀性巴氏杆菌HN07株ompW基因序列(登录号:CP007040.1),使用DNAMAN 5.0软件设计1对引物,选取高保真酶PrimeSTARMax DNA Polymerase进行PCR反应获取目的基因片段,并对ompW基因的核苷酸序列及预测的氨基酸序列进行生物信息学分析。结果表明,PCR扩增产物约为615bp,编码204个氨基酸。核苷酸同源性比对分析显示,羊源多杀性巴氏杆菌ompW基因与猪源、牛源、禽源多杀性巴氏杆菌同源性较高,而与兔源同源性较低。系统进化树结果发现,羊源多杀性巴氏杆菌ompW基因与猪源多杀性巴氏杆菌ompW基因亲缘关系最近。经生物信息学分析发现,ompW蛋白分子式为C1007H1567N257O283S3,分子质量为21.90ku,理论等电点(pI)为9.16,属碱性蛋白质,疏水指数为96.57,总平均疏水性(GRAVY)为0.173(>0),属于疏水类蛋白;前21位氨基酸为信号肽,第5-27位氨基酸区域存在1个跨膜区,存在N-糖基化位点及磷酸化位点,不存在O-糖基化位点,具有多个B细胞、CTL细胞及Th细胞抗原表位;二级结构的α-螺旋、延伸链、β-转角和无规则卷曲分别占17.65%、35.29%、3.92%和43.14%;三级结构是呈β-桶状的单聚体,隶属于外膜蛋白家族成员之一。本研究结果为进一步阐明羊源多杀性巴氏杆菌侵染宿主过程中自身的抗宿主免疫胁迫机制及疫苗的开发与研制提供了理论依据。  相似文献   

9.
为了解四川地区猪环曲病毒(PToV)的发病状况,针对PToV的保守区域S基因设计了一对特异性引物,建立检测PToV的RT-PCR方法,并对其敏感性、特异性和重复性进行了检测。结果显示,仅猪环曲病毒阳性模板可扩增得到约569bp大小的目的条带。敏感性试验显示,该方法能检测到的最低核酸含量为15pg。对采集于12个猪场27个腹泻样本和29个临床健康的样品进行了检测。结果显示,腹泻样本中检出率为33.3%(9/27),而在健康样本中检出率为6.9%(2/29)。以上结果表明本研究成功建立了一种敏感度高、特异性强、重复性良好的用于PToV的RT-PCR方法。  相似文献   

10.
本研究根据猪嵴病毒(porcine kobuvirus,PKV)VP0基因序列特征设计特异性扩增引物,采用RT-PCR方法扩增猪嵴病毒VP0基因全长编码区。将特异性扩增目的片段克隆后进行序列测定,将结果进行拼接后获得猪嵴病毒VP0基因全长并进行相关生物信息学分析。所扩增的目的片段编码有完整的VP0基因开放阅读框,全长为1 098 bp,编码有366个氨基酸,理论等电点为6.71,理论分子质量为38.489 ku,不稳定系数为34.65,最大疏水指数为2.622,最小疏水指数为-2.122。将获得的VP0基因和GenBank中的猪嵴病毒代表株VP0基因序列进行核苷酸同源性比对和遗传进化分析,其与HNXX-4核苷酸同源性最高,为89.1%,与S-1-HUN核苷酸同源性最低,为81.1%。从遗传进化上看,猪嵴病毒VP0基因在遗传进化上呈两个独立的基因亚群,猪嵴病毒中国分离株在两个遗传基因亚群上均有分布。  相似文献   

11.
试验选用96头平均体重14.82 kg左右的杜×长×大断奶仔猪,随机分成4组,每组3栏,每栏8头(公母各半)。对照组饲喂基础日粮,试验1、2、3组分别添加1%80目白术、0.2%白术多糖和1%微米白术。试验期30 d。结果表明:在生长性能方面,与对照组相比,1%微米白术添加组可显著提高日增重(P0.05)、降低饲料增重比和腹泻率,而且效果优于1%80目白术组和0.2%白术多糖组,在肠道形态和肠道微生态区系方面,与对照组相比,日粮添加1%80目白术、0.2%白术多糖、1%微米白术均可不同程度的提高十二指肠和空肠的绒毛高度,加深十二指肠和空肠的隐窝深度,并且增加肠道微生态区系的多样性,其中以1%微米白术添加组的效果最佳。  相似文献   

12.
Sissay, M.M., Uggla, A. and Waller, P.J., XXXX. Prevalence and seasonal incidence of nematode parasites and fluke infections of sheep and goats in eastern Ethiopia. Tropical Animal Health and Production, XXXX. A 2-year abattoir survey was carried out to determine the prevalence, abundance and seasonal incidence of gastro-intestinal (GI) nematodes and trematodes (flukes) of sheep and goats in the semi-arid zone of eastern Ethiopia. During May 2003 to April 2005, viscera including liver, lungs and GI tracts were collected from 655 sheep and 632 goats slaughtered at 4 abattoirs located in the towns of Haramaya, Harar, Dire Dawa and Jijiga in eastern Ethiopia. All animals were raised in the farming areas located within the community boundaries for each town. Collected materials were transported within 24 h to the parasitology laboratory of Haramaya University for immediate processing. Thirteen species belonging to 9 genera of GI nematodes (Haemonchus contortus, Trichostrongylus axei, T. colubriformis, T. vitrinus, Nematodirus filicollis, N. spathiger, Oesophagostomum columbianum, O. venulosum, Strongyloides papillosus, Bunostomum trigonocephalum, Trichuris ovis, Cooperia curticei and Chabertia ovina), and 4 species belonging to 3 genera of trematodes (Fasciola hepatica, F. gigantica, Paramphistomum {Calicohoron} microbothrium and Dicrocoelium dendriticum) were recorded in both sheep and goats. All animals in this investigation were infected with multiple species to varying degrees. The mean burdens of adult nematodes were generally moderate in both sheep and goats and showed patterns of seasonal abundance that corresponded with the bi-modal annual rainfall pattern, with highest burdens around the middle of the rainy season. In both sheep and goats there were significant differences in the mean worm burdens and abundance of the different nematode species between the four geographic locations, with worm burdens in the Haramaya and Harar areas greater than those observed in the Dire Dawa and Jijiga locations. Similar seasonal variations were also observed in the prevalence of flukes. But there were no significant differences in the prevalence of each fluke species between the four locations. Overall, the results showed that Haemonchus, Trichostrongylus, Nematodirus, Oesophagostomum, Fasciola and Paramphistomum species were the most abundant helminth parasites of sheep and goats in eastern Ethiopia.  相似文献   

13.
1-(2-Chloroethyl)3-cyclohexyl-1-nitrosourea (CCNU) is an alkylating agent in the nitrosourea subclass. A prospective evaluation of CCNU was done to determine the maximally tolerated dosage of CCNU in tumor-bearing cats. Response data were obtained when available. Twenty-five cats were treated with CCNU at a dosage of 50-60 mg/m3 body surface area. Complete hematologic data were available for 13 cats. Neutropenia was the acute dose-limiting toxicity. The median neutrophil count at the nadir was 1,000 cells/microL (mean, 2,433 cells/microL; range, 0-9,694 cells/microL). The time of neutrophil nadir was variable, occurring 7-28 days after treatment, and counts sometimes did not return to normal for up to 14 days after the nadir. Based on these findings, a 6-week dosing interval and weekly hematologic monitoring after the 1st treatment with CCNU are recommended. The nadir of the platelet count may occur 14-21 days after treatment. The median platelet count at the nadir was 43,500 cells/microL. No gastrointestinal, renal, or hepatic toxicities were observed after a single CCNU treatment, and additional studies to evaluate the potential for cumulative toxicity should be performed. Five cats with lymphoma and 1 cat with mast cell tumor had measurable responses to CCNU. Phase II studies to evaluate antitumor activity should be completed with a dosing regimen of 50-60 mg/m3 every 6 weeks.  相似文献   

14.
15.
Our particular attention in this article was given to natural mediators for macrophages isolated from the sites of tissue injury. A number of chemotactic factors, which may satisfy many criteria making them acceptable as inflammatory leucocyte chemotactic factors, has been separated. Among them, our laboratory has isolated three macrophage (monocyte) chemotactic factors (MCF-a, -b and -c). Their purification, characterization and functional specificity are discussed.  相似文献   

16.
Human and equine athletes are reported to have a high prevalence of gastric disease, and anecdotal evidence suggests a similar phenomenon applies to racing sled dogs. To investigate the prevalence of gastric disease in racing sled dogs, we conducted 2 gastroscopy studies on dogs competing in the annual Iditarod Sled Dog Race. A pilot study of dogs that were either dropped from the 2000 Iditarod Sled Dog Race because of illness or that finished the race indicated that, approximately 5 days after competing, 10 of 28 dogs (35%) had endoscopic evidence of gastric ulceration, erosion, or hemorrhage. The next year, an endoscopic study of 73 dogs participating in the 2001 Iditarod race was performed in order to evaluate a larger population of dogs. Data from 70 of these dogs could be used; 34 (48.5%) had ulceration, erosion, gastric hemorrhage, or some combination of these findings. When this group of 70 dogs was compared retrospectively to a control group of 87 dogs presented to the Texas A&M University (TAMU) Veterinary Medical Teaching Hospital, the Iditarod sled dogs had a significantly higher prevalence (P = .049) of gastric lesions. These findings suggest that, similar to athletes of other species, elite canine athletes have an increased prevalence of gastric disease compared to the canine population at large.  相似文献   

17.
Genetic variations in chromosome Y are enabling researchers to identify paternal lineages, which are informative for introgressions and migrations. In this study, the male‐specific region markers, sex‐determining region‐Y (SRY), amelogenin (AMELY) and zinc finger (ZFY) were analysed in seven Turkish native goat breeds, Angora, Kilis, Hair, Honaml?, Norduz, Gürcü and Abaza. A SNP in the ZFY gene defined a new haplotype Y2C. All domestic haplogroups originate from Capra aegagrus, while the finding of Y1A, Y1B, Y2A and Y2C in 32, 4, 126 and 2 Turkish domestic goats, respectively, appears to indicate a predomestic origin of the major haplotypes. The occurrence of four haplotypes in the Hair goat and, in contrast, a frequency of 96% of Y1A in the Kilis breed illustrate that Y‐chromosomal variants have a more breed‐dependent distribution than mitochondrial or autosomal DNA. This probably reflects male founder effects, but a role in adaptation cannot be excluded.  相似文献   

18.
Continuous rate infusion (CRI) of furosemide in humans is considered superior to intermittent administration (IA). This study examined whether furosemide CRI, compared with IA, would increase diuretic efficacy with decreased fluid and electrolyte fluctuations and activation of the renin-angiotensin-aldosterone system (RAAS) in the horse. Five mares were used in a crossover-design study. During a 24-hour period, each horse received a total of 3 mg/kg furosemide by either CRI (0.12 mg/kg/h preceded by a loading dose of 0.12 mg/kg IV) or IA (1 mg/kg IV q8h). There was not a statistically significant difference in urine volume over 24 hours between methods; however, urine volume was significantly greater after CRI compared with IA during the first 8 hours ([median 25th percentile, 75th percentile]: 9.6 L [8.9, 14.4] for CRI versus 5.9 L [5.3, 6.0] for IA). CRI produced a more uniform urine flow, decreased fluctuations in plasma volume, and suppressed renal concentrating ability throughout the infusion period. Potassium, Ca, and Cl excretion was greater during CRI than IA (1,133 mmol [1.110, 1,229] versus 764 mmol [709, 904], 102.7 mmol [96.0, 117.2] versus 73.3 mmol [65.0, 73.5], and 1,776 mmol [1,657, 2.378] versus 1,596 mmol [1,457, 1,767], respectively). Elimination half-lives of furosemide were 1.35 and 0.47 hours for CRI and IA, respectively. The area under the excretion rate curve was 1,285.7 and 184.2 mL x mg/mL for CRI and IA, respectively. Furosemide CRI (0.12 mg/kg/h) for 8 hours, preceded by a loading dose (0.12 mg/kg), is recommended when profound diuresis is needed acutely in horses.  相似文献   

19.
The objective of this report was to characterize 20-year changes in proportion of calcium oxalate (CaOx) calculi and struvite calculi in dogs, and associations with breed, age, and sex. In this retrospective study, results of analysis of urinary calculi from dogs were reviewed for specimens received between July 1, 1981, and December 31, 2001. Breed, sex, age, year of submission of the specimen, and mineral type(s) were analyzed statistically. CaOx or Struvite or both were contained in 18,966 of 20,884 (91%) specimens. For both sexes, a 20-year statistically significant increase was observed in the proportion of calculus specimens that contained CaOx. The increase in this proportion was greater in females (1% to 31%) than in males (18% to 82%). From 1998 to 2001, when proportions may have plateaued, the odds of specimens containing CaOx were markedly higher in 18 breeds, markedly lower in 5 breeds, and not significantly different in 13 breeds compared with crossbreds. For both sexes, a 20-year statistically significant decrease was observed in the proportion of calculus specimens that contained struvite. This decrease in proportion was greater for males (79-16%) than for females (97-68%). From 1998 to 2001, when proportions plateaued, the odds of calculi containing struvite were markedly lower in 20 breeds, markedly higher in 1 breed, and not significantly different in 15 breeds when compared with crossbreds. Breed, age, and sex were associated statistically with CaOx or struvite urolithiasis. In conclusion, there appears to have been a long-term increase in the proportion of specimens of canine urinary calculi that contain CaOx as well as a long-term decrease in the proportion of specimens of calculi that contain struvite for both male and female dogs. The rate of change appeared to begin leveling off in the period 1998 to 2001. The recent proportion of dogs with either CaOx- or struvite-associated urolithiasis may depend on breed, age, and sex, and on interactions among these 3 factors.  相似文献   

20.
The epidemiology of H. placei and of other gastrointestinal nematodes in yearling dairy cattle was examined on two farms in Kiambu District, central Kenya during each of 13 one-month periods from April 1993 to April 1994. On each farm, 32 newly weaned dairy calves were given a single dose of albendazole and then placed on experimental pastures. Twelve of the animals were designated for bi-monthly slaughter (n = 2) and analysis of worm population characteristics and 20 were designated for blood and faecal collection and for weighing. Two parasite-free tracer calves were grazed alongside the weaner calves each month throughout the study period and were also slaughtered for analysis of worm populations. Faecal egg counts, haematological and serum pepsinogen determinations, herbage larval counts, and animal live weight changes were recorded monthly. The study revealed that Haemonchus placei, Trichostrongylus axei, Cooperia spp. and Oesophagostomum radiatum were responsible for parasitic gastroenteritis and that H. placei was the predominant nematode present in the young cattle on both farms. Faecal egg counts from resident cattle and necropsy worm counts revealed that pasture larval levels were directly related to the amount of rainfall. The total worm burdens in the animals were highest during the rainy season (March–June and October–December) and lowest during the dry seasons (July–September and January–February). The very low recovery of immature larvae of H. placei from the tracer calves indicated that arrested development is not a feature of the life cycle of this parasite in central Kenya. The maintenance of the parasite population depended on continuous cycling of infection between the host and the pasture. The agroclimatic conditions of the study area were such that, in general, favourable weather conditions for the development and survival of the free-living stages of gastrointestinal nematodes existed all year round.  相似文献   

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