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1.
杜大长猪IGF-I基因的克隆及其序列分析   总被引:5,自引:0,他引:5  
采用TRIzol法从杜大长杂交猪的肝脏中提出总RNA,将其纯化后作为PCR扩增模板 ,以设计的P1(5′-CTACATTCTGTAGTTCTTGTTTCC-3′)为引物合成IGF-I基因cDNA的第1链,再以P1和P2(5′-ATGGCCCTGTGCTTGCTCTCCTT-3′)为引物扩增到大小约为360bp的产和,并将其达隆至pGEM-T载体上。经筛选、酶切、我分析,表明该片段为IGF-I基因的cDNA  相似文献   

2.
本试验根据Lamb等[1]报道的鸡生长激素基因序列,设计合成2对特异性引物PM3和PMPS1,它们的碱基序列分别是PM3FOR5′-atccccaggcaaacatcctc-3′REV5′-cctcgacatccagctcacat-3′PMPS1FOR5′-ctaaaggacctggaagaaggg-3′REV5′-aacttgtcgtaggtgggtctg-3′。通过PCR反应从鸡基因组中扩增出含有GH基因内含子1和4的片段,再用MSP1内切酶对产物做RFLPs分析。由于特异性PCR的条件要…  相似文献   

3.
三品系西方蜜蜂基因组DNA多态性分析研究(二)   总被引:2,自引:0,他引:2  
采用9种随机引物(K,5’-CGGCCCCTGT-3’;5’-CAGCGGTACC-3’;5’-ACCAGGTTGG-3’,5’-GTCGGAATTC-3’;5’-CGGCCCCGGT-3’;X,5’-GTAGTCACAC-3’;Y,5’-CTGCAGGACA-3’;Z,5’-CGGCCCGGTA-3’)和PCR技术扩增不同产蜜量的三品系西峰(喀尼阿兰、平湖、美意的DNA基因组。结果K引物在高产蜜  相似文献   

4.
具有中和活性的减蛋综合征病毒单抗的研制   总被引:1,自引:0,他引:1  
用减蛋综合征病毒毒株WPDV205纯化抗原免疫BALB/c小鼠,在最后一次免疫后第3天取脾细胞与SP2/0细胞在PEG-4000的作用下融合。用间接ELISA初步筛选出阳性克隆10株(4B1、1D4、2D6、3D6、3A5、2A1、3C6、3C1、1B3和2C5)。这些杂交瘤细胞经克隆化和再次检测后生产腹水,用微量细胞中和试验筛选出具有中和活性的单克抗隆体4株(4B1、3C1、3C6和2C5),其  相似文献   

5.
4型菌毛、毒素及蛋白酶在动物源气单胞菌的检测   总被引:1,自引:0,他引:1  
将来源于鱼、猪、犬等9种动物的18株气单胞菌分离株通过AD(氨苄青霉素-糊精)培养基纯化。另以大肠杆菌束状菌毛相关(bfp-related)基因的PCR引物对EB1(GATTGAATCTGCAATGGTGC)—EB2(GGATTACTGTCCTCACATAT)扩增,用digoxingenin-11-dUTP标记,构建核酸探针,作斑点杂交试验,用以检测上述气单胞菌菌株的4型菌毛,同时以溶血试验检测其毒素,用脱脂奶溶蛋白圈试验检测其蛋白酶。结果显示,在18个菌株中,4型菌毛、毒素和蛋白酶均为阳性者为10株(55.6%);3项指标均无肯定的阳性结果者4株,两者合计共14株,占检测总数约80%(14/18),提示这3种毒力因子存在较高的相关性。所有鱼源株(5株)三项指标皆为阳性;4株猪分离株三项指标均为阳性者占一半;其它动物分离株检测结果无明显规律性  相似文献   

6.
基因探针和PCR对鲤吉陶单极虫的检测   总被引:4,自引:1,他引:3  
采用液氮冷冻研磨法破碎吉陶单极虫(Thelohaneluskitauei)孢子,按常规法提取其基因组DNA。取DNAEcoRI消化产物,采用低熔点琼脂糖回收法制备大(3.0~15.0kb)、小(0.5~3.0kb)片段,将其克隆于pBluescriptksEcoRI位点,经α-互补现象、酶切鉴定和虫体DNA生物素标记探针筛选,构建了含23个克隆的基因文库,克隆片段介于0.9~6.8kb之间;经阳性克隆标记筛选及特性分析,制备了长度为0.9kb(19号质粒克隆片段)的探针,该探针具有较强的敏感性和特异性。对该探针两端DNA序列进行分析,设计出1对引物。上游引物序列为:5′TTTCGAACCCGCACAACAAG3′,下游引物序列为:5′TGAGTGGATAG-TATCGCTGC3′。应用该对引物对虫体进行了检测  相似文献   

7.
经SephadexG75和PAGE分离,以双波长薄层自动扫描仪扫描测定了家兔胚源性特异蛋白-1(rESP-1)的分泌特性;离体组织培养分析了rESP-1对子宫蛋白质合成的影响。结果表明,早孕期rESP-1的分泌呈二次曲线,第3天分泌出现,第4天出现第一峰值,第5天下降,然后迅速增加,最大分泌在第7天,第8天迅速回降。其分泌量随胚胎数(或黄体数)的增加而升高,且受FSH-LH和PMSG-LH的影响。rESP-1使子宫细胞RNA增加,促进子宫蛋白质合成  相似文献   

8.
用EooRI+HindⅢ双酶解含鸡α-珠蛋白基因5′端核基质附着区(MAR)、小鼠金属硫蛋白基因启动子(MT-1)和人胰岛素样生长因子1-(hIGF-1)基因的pMTSMCAG质粒,0.6%琼脂糖凝胶电泳,玻璃乳回收纯化MAR/MT/hIGF-1片段(3.9kb),用该片段制备转基因兔。利用水平显微注射系统注射1-细胞兔胚胎468枚,移植到36中受体兔,获得8只阳性兔(显示出约600bp外源DNA  相似文献   

9.
用EcoRI+HindⅢ双酶解含鸡α-珠蛋白基因5′端核基质附着区(MAR)、小鼠金属硫蛋白基因启动子(MT-1)和人胰岛素样生长因子-1(hIGF-1)基因的pMTSMCAG质粒,0.6%琼脂糖凝胶电泳,玻璃乳回收纯化MAR/MT/hIGF-1片段(3.9kb),用该片段制备转基因兔。利用水平显微注射系统注射1-细胞兔胚胎468枚,移植到36只受体兔,有8只妊娠,共产下19只活仔兔。采仔兔耳样提取基因组DNA,应用PCR技术分析其外源基因整合情况,获得8只阳性兔(显示出约600bp外源DNA条带)。再对8只阳性兔的PCR产物进行Southern印迹杂交,得到5只整合有MAR/MT/hIGF-1外源基因的转基因兔,转基因整合率为26.3%(5/19)。用1只阳性转基因公兔(502号)繁殖了6窝,共获得42只仔兔。采仔兔耳样提取基因组DNA后,如上用PCR分析和做Southern印迹杂交,F1代中有5只整合有MAR/MT/hIGF-1融合基因。  相似文献   

10.
鸡传染性支气管炎病毒S1基因在昆虫细胞中的高效表达   总被引:8,自引:1,他引:7  
将鸡传染性支气管炎病毒( I B V) S1 基因 c D N A 克隆至含有起始密码 A T G 的转移载体 p S X I V V I+ X3/4,构建成重组转移质粒 p S X I V V I+ X3/4 S1. Holte,再与粉纹夜蛾核型多角体病毒 Tn N P V S V I- G D N A( O C C- , gal+ )共转染 草地夜蛾( Sf9) 细胞, 经空斑纯化得 到已插入 S1 基 因并能形成多角体 的重组病毒 Tn N P V( X3/4) S1. Holte O C C+ 。以重组毒株感染 Tn5 B1 细胞,在不同时间收取感染了病毒的细胞进行 S D S P A G E 与 W estern 印迹检测。 Tn N P V( X3/4) S1. Holte O C C+ 在感染的细胞中高效表达了 S1 基因,表达产物为约 100 000 的融合蛋白,与预计的表达修饰后的产物大小相符,推测此蛋白已糖基化。 S D S P A G E凝胶薄层色谱分析结果显示,感染病毒后 48、72、96 h, S1 蛋白分别占细胞内总蛋白量的 287% 、358% 、371% 。  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.
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   

15.
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   

16.
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   

17.
The purpose of this study was to determine the frequency of different tumor types within a large cohort of cats with intracranial neoplasia and to attempt to correlate signalment, tumor size and location, and survival time for each tumor. Medical records of 160 cats with confirmed intracranial neoplasia evaluated between 1985 and 2001 were reviewed. Parameters evaluated included age, sex, breed, FeLV/FIV status, clinical signs, duration of signs, number of tumors, tumor location(s), imaging results, treatment, survival times, and histopathologic diagnosis. Most of the cats were older (11.3 +/- 3.8 years). Primary tumors accounted for 70.6% of cases. Metastasis and direct extension of secondary tumors accounted for only 5.6 and 3.8% of cases, respectively. Twelve cats (7.5%) had 2 or more discrete tumors of the same type, whereas 16 cats (10.0%) had 2 different types of intracranial tumors. The most common tumor types were meningioma (n = 93, 58.1%), lymphoma (n = 23, 14.4%), pituitary tumors (n = 14, 8.8%), and gliomas (n = 12, 7.5%). The most common neurological signs were altered consciousness (n = 42, 26.2%), circling (n = 36, 22.5%), and seizures (n = 36, 22.5%). Cats without specific neurological signs were common (n = 34, 21.2%). The tumor was considered an incidental finding in 30 (18.8%) cats. In addition to expected relationships (eg, meninges and meningioma, pituitary and pituitary tumors), we found that lesion location was predictive of tumor type with diffuse cerebral or brainstem involvement predictive of lymphoma and third ventricle involvement predictive of meningioma.  相似文献   

18.
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   

19.
The occurrence of ectoparasites in sheep flocks is frequently reported but seldom quantified. Sheep production used to be a predominantly family activity in the state of Sa~o Paulo (Brazil), but it began to become a commercial activity in the past decade. Thus, information about the ectoparasites existing in sheep flocks has become necessary. The present data were obtained by means of questionnaires sent to all sheep breeders belonging to the `Associaça~o Paulista de Criadores de Ovinos' (ASPACO; Sa~o Paulo State Association of Sheep Breeders). Response reliability was tested by means of random visits paid to 10.6% of the respondents. Most of the properties (89.5%) reported the presence of one or more ectoparasites. Screw-worm (Cochliomyia hominivorax) was the most frequent ectoparasite (72.5%), followed by bot fly larvae (Dermatobia hominis, 45.0%), ticks (Amblyomma cajennense) and Boophilus microplus, 31.3%) and finally lice (Damalinia ovis, 13.8%). Combined infestations also occurred, the most common one being screw-worm with bot fly larvae (36.0%) followed by bot fly larvae with ticks (13.9%), screw-worm with ticks (9.3%), bot fly larvae with lice (6.9%), and ticks with lice (5.0%). The most common triple combination was screw-worm, bot fly larvae and ticks (12.8%). Breeds raised for meat or wool were attacked by bot fly larvae and ticks more often than other breeds. Lice were only absent from animals of indigenous breeds. The relationships among these ectoparasites are discussed in terms of sheep breeds, flock size, seasonality and the ectoparasitic combinations on the host.  相似文献   

20.
Infectious diseases have always been a terrible scourge for humans. The appearance of these plagues, as they were called without distinction, was generally connected to various conditions: asters, climatic changes or religious reasons. The concept of contagious, and then infectious, diseases came slowly. Variolation, i.e. transmission of ‘virulent’ matter to induce a natural disease and the immunity against it, was brought from Constantinople to England by Lady Montague, in 1721. This ‘variolation’ technique was also often performed in veterinary medicine against diseases like sheep-pox or pleuropneumonia. As ‘vaccination’ is the term generally accepted for ‘immunisation’, variolation can be the word designating such a technique. The second period of the history of immunisation began, in 1880, with the studies of Pasteur and his collaborators. A great number of bacterial vaccines were developed: dead, live but attenuated or only parts of pathogens. The viruses were produced in animals, then in eggs and at last, in tissue cultures. Second generation vaccines appeared with genetic engineering: recombinant vaccines, vector vaccines, nucleic acids vaccines, and markers vaccines, among others. These novel technologies can permit the development of new ones and improve the quality of the vaccines already existing.  相似文献   

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