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1.
本试验旨在克隆并表达中国荷斯坦奶牛髓样分化蛋白-2(myeloid differentiation protein -2,MD-2)基因,并用Western blotting进行鉴定。采用RT-PCR技术从中国荷斯坦奶牛外周血白细胞中扩增MD-2 cDNA,并将扩增产物与pMD20-T载体连接,重组质粒经测序鉴定后,构建pET28a-MD-2重组质粒,转化大肠杆菌BL21(DE3),经IPTG诱导表达后进行SDS-PAGE和Western blotting分析。结果表明,目的基因含有1个483 bp的开放阅读框,编码161个氨基酸;与黄牛、黑猩猩和小鼠MD-2的cDNA序列同源性分别为99.38%、77.02%、68.12%,氨基酸同源性分别为98.75%、65.00%和58.13%。融合蛋白以包涵体的形式存在,分子质量约为25 ku。说明本研究成功克隆并表达了中国荷斯坦奶牛MD-2 cDNA,为其进一步的功能研究提供了理论依据。  相似文献   

2.
本试验旨在扩增版纳微型猪近交系DLDH基因编码区序列,为进一步研究DLDH基因的表达信号通路及与精子获能和顶体反应发生相关基因的表达调控提供必要的基础。通过提取版纳微型猪近交系睾丸组织RNA,经反转录后获得cDNA,设计特异性引物以扩增DLDH基因的编码区序列,经测序后采用生物信息学软件对编码区序列及氨基酸序列进行生物信息学分析和结构预测。结果表明,版纳微型猪近交系DLDH基因编码区序列全长1 530bp,编码509个氨基酸。生物信息学分析表明,不同物种DLDH基因编码区核苷酸序列和氨基酸序列具有较强的保守性,DLDH蛋白中存在丰富的磷酸化位点,空间结构以α-螺旋和无规则卷曲为主。  相似文献   

3.
试验旨在克隆中国荷斯坦奶牛CC趋化因子受体5(C-C chemokine receptor type 5,CCR5)基因,对其进行生物信息学分析,并探究CCR5基因在奶牛炎性和健康组织中的表达水平。采用PCR技术扩增并克隆荷斯坦奶牛CCR5基因CDS区全长序列,连接pMD18-T载体并转化大肠杆菌DH5α感受态细胞,通过蓝白斑方法筛选阳性克隆并测序,对序列进行相似性比对及系统进化树构建;应用多种在线生物信息学软件对其编码蛋白进行分析,并利用实时荧光定量PCR方法检测CCR5基因在健康和炎症奶牛乳腺组织中的表达情况。结果显示,中国荷斯坦奶牛CCR5基因CDS区全长1 059 bp,编码352个氨基酸。相似性和遗传进化分析结果显示,奶牛CCR5基因与绵羊的遗传距离最近,高达96.0%,与鸡遗传关系最远,为61.0%,且在不同物种之间CCR5基因高度保守。中国荷斯坦奶牛CCR5蛋白分子质量为40.235 ku,理论等电点(pI)为9.30,为疏水性蛋白但不是分泌蛋白,主要存在于细胞质内;在CCR5蛋白二级结构中α-螺旋和无规则卷曲分别占51.14%和32.95%,三级结构模型预测结果与二级结构一致。实时荧光定量PCR结果显示,CCR5基因在健康奶牛乳腺组织中的表达量极显著低于炎性奶牛乳腺组织(P<0.01),提示其可能参与奶牛乳腺炎的发生过程。本试验结果为进一步研究奶牛CCR5蛋白的功能提供了理论依据,对探究奶牛CCR5基因在奶牛乳腺炎中的调控功能等具有重要意义。  相似文献   

4.
为了进一步了解鹿源DCN基因的结构与功能,揭示该基因在鹿茸尖端不同组织层的表达规律,本研究从梅花鹿鹿茸尖端组织cDNA文库中首次克隆具有完整编码区的DCN基因全长cDNA序列,并结合生物信息学方法和实时荧光定量RT-PCR技术对该基因的氨基酸序列结构和表达特征进行分析.结果表明,梅花鹿DCN基因cDNA全长为1 831 bp,编码360个氨基酸;其编码蛋白具有N端信号肽,相对分子质量为39.9 ku,理论等电点为8.8,其一级结构中亮氨酸所占比例最高(12.5%);梅花鹿与绵羊DCN氨基酸序列的相似性最高(98%).实时荧光定量RT-PCR分析表明,DCN在间充质层的表达显著高于其他3层组织,提示DCN的抗纤维化活性可能是维持鹿茸间充质层快速生长的重要调节因素.  相似文献   

5.
从梅花鹿鹿茸尖端组织全长cDNA文库中克隆了与骨形成和骨改建有关的一种新的骨生长因子BSPⅡ基因的全长cDNA序列,并结合生物信息学方法和实时荧光定量RT-PCR技术对该基因的氨基酸序列及其在鹿茸尖端不同组织层的表达情况进行了分析。结果表明,BSPⅡ基因cDNA全长为1576bp,编码311个氨基酸。经生物信息学分析表明,该基因编码的蛋白具有N端信号肽及跨膜区,相对分子质量为34100,理论等电点为4．05,其一级结构中谷氨酸所占比例最高;二级结构元件主要以α-螺旋和无规则卷曲为主;同源序列分析表明,梅花鹿与欧洲牛BSPⅡ氨基酸的相似性最高（93％）;多重序列比对显示,此蛋白的N-端和68～215、265～308位谷氨酸富集区高度保守;系统进化树显示,在该基因座位上梅花鹿与马的亲缘关系较近,来源于同一个分化支。实时荧光定量RT—PCR分析表明,该基因的表达与鹿茸组织的矿化进程呈显著正相关,推测该基因在鹿茸组织矿化过程中起到了重要的调节作用。  相似文献   

6.
为了获得荷斯坦牛DQA2基因序列并研究其序列特征及编码蛋白的结构和功能,本研究采用DNA混合池扩增后直接测序的方法,通过DNAstar软件和在线服务器预测中国荷斯坦牛DQA2基因CDS区编码蛋白的理化性质、疏水性/亲水性、糖基化位点、信号肽、跨膜结构、二级结构以及三级结构。结果表明,中国荷斯坦牛DQA2基因CDS区长度为765bp,编码254个氨基酸,蛋白结构预测是一种混合型的可溶性蛋白质,具有4个潜在的N-糖基化位点,分别为Asn31、Asn96、Asn260、Asn369;在第1~23位氨基酸之间存在信号肽;存在1个典型的跨膜螺旋区,氨基酸序号为217-239。本研究结果为进一步研究荷斯坦牛的抗病育种提供了理论基础。  相似文献   

7.
柳蚕(Actias selene Hübner)是鳞翅目大蚕蛾科的一种珍稀野生绢丝昆虫.利用RT-PCR方法克隆了柳蚕丝素重链基因(Fib-H)cDNA 5'端的一个片段,该片段序列长819 bp, 编码273个氨基酸,含有4个保守多聚丙氨酸结构域.序列比对分析表明,该片段序列与樟蚕(Saturnia japonica)、天蚕(Antheraea yamamai)、柞蚕(Antheraea pernyi)、印度柞蚕(Antheraea mylitta)和家蚕(Bombyx mori) 的Fib-H基因cDNA 5'端同源序列的相似性分别为72.5%、65.8%、65.1%、65.1%、47.1%.半定量PCR分析显示柳蚕Fib-H基因cDNA 5'端片段序列在5龄幼虫第1-12天的表达量呈现逐渐上升的趋势.  相似文献   

8.
以奶牛骨组织提取的RNA为模板，利用RT—PCR技术扩增出奶牛骨钙素全长cDNA，然后将扩增产物重组到PMD-18T载体中，测定了全基因的核苷酸序列。序列分析表明，奶牛骨钙素全长cDNA为303bp，编码100个氨基酸，与GenBank中的X53699的序列完全相同。通过加端PCR技术连接单链DNA片段人工定点同义突变，将奶牛骨钙素成熟蛋白基因中的大肠杆菌稀有密码子同义突变为大肠杆菌常用密码子并亚克隆至PET-32a表达载体．转化到宿主菌BL21（DE3）中，经IPTG诱导后，成功表达出奶牛骨钙素融合蛋白。  相似文献   

9.
为鉴定牦牛IFITM1基因并分析其在不同生长阶段牦牛5种组织中的转录水平和蛋白表达量.本研究采用根据普通牛IFITM1基因设计的引物,PCR扩增牦牛IFITM1基因并利用生物信息学软件分析其序列,结果显示,获得牦牛546 bp的IFITM1基因cDNA序列,其中编码(CDS)区为375 bp,编码124个氨基酸残基.牦...  相似文献   

10.
奶牛朊病毒基因克隆与序列分析   总被引：8，自引：0，他引：8  
根据已报道正常牛朊蛋白(PrP^c)基因(PRNP)序列设计引物，采用PCR法扩增了6头荷斯坦奶牛的PRNP基因，将其克隆到T-Vector。序列测定及分析表明所克隆的奶牛PRNP基因片段为795bp，该基因内无内含子，包含了牛PRNP完整编码区序列，编码264个氨基酸的前体蛋白，推测其分子量约34ku。其中2头共同含有未曾报道的牛PRNP多态性位点M120I，无义突变G234A，但未引起酶切位点变异，未发现插入或缺失变异；与已报道牛PRNP序列(GenBank收录号为DI0613)相比，两者核苷酸序列同源性为99％，其编码的氨基酸同源性为99％。  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.
Tropical Animal Health and Production -  相似文献   

14.
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   

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.
<正>1.INTRODUCTION Ensuring transportation network security is one Of the most daunting challenges confronting homeland security agencies today.Significant research has been dedicated.To model and analyze the vulnerability of transportation systems,while notably fewer studies propose specific strategies for deploying defensive technologies to safeguard these systems.  相似文献   

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