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1.
根据GenBank中旋毛虫53000抗原基因的序列设计引物，用PCR技术直接从旋毛虫肌幼虫cDNA文库中扩增靶基因p53cDNA，将其克隆到pMD-18T载体，转化至大肠埃希氏菌NovaBlue后测序分析，结果表明，克隆到1176bp的p53cDNA，共编码391个氨基酸。序列分析表明，p53cDNA序列与GenBank中的旋毛虫p53基因序列相比共有12个核苷酸发生改变，二者的同源性为98％，所编码蛋白质氨基酸序列的同源性为98％。将其克隆到原核表达载体pET28a并转化至表达菌BL21star（DE3），经IPTG诱导表达后获得约48000的重组蛋白。Western blotting检测证实，p53重组蛋白可以被旋毛虫感染猪血清识别，具有很好的抗原性。 相似文献
2.
对狂犬病病毒（RV）ERA株核蛋白（N）基因进行了克隆和序列分析。根据GenBank中已发表的RVN基因序列，设计合成了1对特异性引物，对RV ERA株N基因进行了RT-PCR扩增。将PCR产物纯化后与pMD18T连接得到重组质粒pMD-N，并进行核苷酸序列测定。结果该基因全长1353bp，编码450个氨基酸。RV ERA株与CVS-11、PV-11、SRV9和SAD—B19株相比，核苷酸的同源性分别为98％、98．3％、99％、99.6％，推导的氨基酸的同源性分别为98％、99％、99％、99.6％。将pMD-N双酶切，回收目的基因片段并克隆到原核表达载体pET28a（＋）中，构建了重组质粒pETN；将其转化表达菌BL21（DE3）并用IPTG进行诱导表达。结果重组菌裂解物经SDS-PAGE电泳可检测到相对分子量为53kDa的重组蛋白。经凝胶薄层扫描分析，重组蛋白表达量占菌体蛋白的56．8％，Western-blot结果显示该重组蛋白能与RV多克隆抗体发生特异性反应。 相似文献
3.
以胸膜肺炎放线杆菌（APP）血清1型标准菌株基因组为模板，用PCR扩增ApxⅠ BD基因的特异性片段；对PCR产物纯化回收后与载体pMD18-T进行连接、转化，经酶切和序列分析鉴定后亚克隆到原核表达载体pET28a中，构建重组表达质粒pETIBD；转入大肠埃希氏菌BL21中，以IPTG诱导表达，进行SDS-PAGE电泳。结果，从APP中克隆的ApxⅠ BDDNA序列与已发表序列的同源性为100％，长1447bp，编码482个氨基酸，所表达的融合蛋白相对分子质量约54ku，与预期的大小相符。结果表明，含有ApxⅠBD基因特异性片段的重组表达载体pETⅠBD已成功构建。 相似文献
4.
从江香猪IFN-β基因的序列分析及原核表达 总被引：1，自引：1，他引：0
试验旨在探明从江香猪β-干扰素（interferon-beta，IFN-β）基因编码区分子序列及原核表达产物特征。以从江香猪为研究对象，提取肝脏总RNA并反转录为cDNA，设计特异性引物扩增IFN-β基因编码区，将目的基因片段克隆至原核表达质粒pET-28a上，获得重组质粒pET28a-CJpoIFN-β，并利用生物学软件对江香猪IFN-β基因编码区进行序列分析；将鉴定正确的重组质粒pET28a-CJpoIFN-β转化大肠杆菌BL21（DE3）感受态细胞，经IPTG诱导表达、SDS-PAGE与Western blotting分析原核表达蛋白。结果表明，从江香猪IFN-β基因编码区长为561 bp，编码186个氨基酸；该蛋白为分泌性蛋白，前21个氨基酸为信号肽序列；二级结构主要以α-螺旋（77.42%）和无规则卷曲（17.74%）为主。从江香猪与其他猪源IFN-β基因核苷酸序列同源性为99.5%~100.0%，与禽的同源性最低（35.2%）；从江香猪与巴马猪、梅山猪IFN-β氨基酸同源性均为100.0%，但与贵州白香猪IFN-β同源性为99.5%，存在E43Q、K73R和C161R 3处氨基酸的差异。Western blotting结果显示，带His标签的重组表达蛋白能被His单抗识别，条带大小约为24 ku。本试验结果为进一步研究IFN-β基因生物学活性及加快从江香猪这一品种资源的有效利用提供参考依据。 相似文献
5.
根据巴氏杆菌磺胺耐药基因Sul Ⅱ序列设计引物，以临床分离的禽源巴氏杆菌磺胺耐药菌株为模板，扩增出禽源巴氏杆菌Sul Ⅱ的全长基因序列。经T载体克隆和核苷酸序列测定，克隆的Sul Ⅱ基因与文献报道的同源性为98．9%，有9个碱基的差异，但所编码的氨基酸没有改变。将Sul Ⅱ基因按正向的阅读框架与表达戴体pET-28c( )连接，重组质粒经酶切鉴定正确后，经IPTG诱导，SDS-PAGE检测，融合蛋白的表达产物占菌体总蛋白的11％，蛋白表达形式为包涵体表达。 相似文献
6.
为了探索牛乳腺炎无乳链球菌磷酸甘油酸激酶（Phosphoglycerate kinase,Pgk）基因编码蛋白的抗原性,根据GenBank公布的牛源无乳链球菌Pgk基因序列,设计并合成一对引物,通过PCR扩增出Pgk蛋白抗原优势区的编码序列,并对其进行克隆、测序、转化、诱导表达及抗原性鉴定。结果显示克隆的Pgk基因序列含有984 bp,编码328个氨基酸残基。临床分离株与GenBank上公布的B群无乳链球菌菌株Pgk基因（AE009948）核苷酸序列同源性为99.09%,氨基酸序列同源性为99.69%。将扩增的Pgk基因片段克隆至原核表达载体pET30a（＋）中,构建重组表达载体Pgk-pET30a（＋）,再将筛选的阳性重组质粒转化至BL21工程菌,经IPTG诱导获得部分可溶性表达Pgk重组蛋白。经Ni2＋亲和层析柱纯化得到纯度在90%以上的蛋白,经Western blot分析结果表明,重组蛋白具有良好的抗原性,为进一步探索其对奶牛的免疫原性研究奠定了良好的实验基础。 相似文献
7.
鹅副粘病毒NP基因的原核表达及表达产物抗原性检测 总被引：1，自引：0，他引：1
根据GenBank中发表的GPMVSF02株的NP基因的核苷酸序列设计合成一对引物，通过RT-PCR扩增JS／1／97／Go株的NP基因，将其克隆人pMDl8-T载体，序列测定和分析表明：所扩增的NP基因的核苷酸长为1470bp，共编码490个氨基酸。再将NP基因定向亚克隆至原核表达载体pGEX-6P的多克隆位点，经酶切鉴定后将含有NP基因的阳性亚克隆重组质粒转化到大肠杆菌BL21中，用诱导荆IPTG分别以不同的浓度进行诱导，并在不同的诱导时间进行采样，样品经处理后通过SDS-PAGE、Westernblot和Dot-ELISA进行分析。结果显示：以终浓度为1mmol／LIPTG进行诱导4h表达可达到高峰。表达的融合蛋白大小约为80kD，并具有良好的抗原性。 相似文献
8.
采用PCR方法扩增驽巴贝虫吉林分离株BC-48基因片段，将扩增产砌与pGEM—TEasy载体连接，重组质粒经PCR、单酶切鉴定后测序；构建BC-48的重组pGEX-4T-2表达载体．经IPTG诱导表达后，进行SDS-PAGE、Western—blotting分析。结果显示，克隆的BC-48基因片段长610bp，含有一个570bp的开放阅读框，编码189个氨基酸，与GenBank中USDA株（U46551）的同源性为96．7％；表达的融合蛋白为45ku，能被驽巴贝虫阳性血清识别；表明该融合蛋白具有较好的反应原性。 相似文献
9.
家蚕细胞色素P450 CYP337A1的原核表达 总被引：1，自引：0，他引：1
以含有CYP337A1基因完全编码框的克隆载体质粒DNA为模板进行PCR扩增,产物经TA克隆测序鉴定后,亚克隆至原核融合表达载体PET50（b）（含有编码64．3kD的Nus．Tag蛋白标签）中,在大肠杆菌BL21（DE3）中进行原核表达。利用SDS-PAGE方法,以空载和未诱导菌体作对照,对细菌总蛋白进行电泳分析,结果表明：经1mmol／L IPTG诱导2h后,即能看到1条大约120kD的蛋白质特异带,与预计大小一致。进一步利用不同时间、不同温度以及不同浓度的IPTG进行诱导结果比较,发现在诱导时间为2h,温度为32℃,IPTG浓度为0．6mmol／L的诱导条件下,融合蛋白表达量最大。该实验为利用该基因的蛋白产物进行功能分析创造了条件。 相似文献
10.
根据旋毛虫新生幼虫期特异性基因pBK—CMV—N10序列设计引物，利用PCR技术将基因N10的信号肤去除后，用T／A法克隆到pMD-18T载体，转化至大肠杆菌NovaBlue，经鉴定及序列测定，结果显示成功克隆到N10基因。将重组质粒pMD-18T—N10进行酶切后连接到原核表述载体pET-28a中，重组质粒经鉴定后转化大肠杆菌BL21star（DES），用IPTG诱导表达出与理论相符的融合蛋白，相对分子质量为38700，诱导4h的表达量占菌体总蛋白量的15％。从N10重组蛋白提取可溶性融合蛋白，对其生物学特性初步鉴定结果表明，具有类似脱氧核糖核酸酶Ⅱ的活性。 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.
B. Cinar Kul N. Bilgen J.A. Lenstra O. Korkmaz Agaoglu B. Akyuz O. Ertugrul 《Zeitschrift für Tierzüchtung und Züchtungsbiologie》2015,132(6):449-453
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
14.
Bazin H 《Comparative immunology, microbiology and infectious diseases》2003,26(5-6):293-308
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
15.
The ash, silica and certain important micronutrients were estimated in conventional and unconventional feed and fodder resources available in Southern India. Commonly used dry roughages, such as paddy straw, ragi straw, maize kadbi/stalk, jowar kadbi/stalk, bajra stalk and wheat straw, were high in ash (9.9% +/- 0.77%) and silica (6.4% +/- 0.65%) and low in most of the other micronutrients, except iron, with paddy straw containing most silica (>9%). Cultivated non-leguminous (maize, jowar) and improved green crops (hybrid napier, guinea, green panic, NB-21, CO-1) were also high in ash (10.5% +/- 0.60% and 12.5% +/- 0.51%) but were moderate sources of P, Mg and Cu and good sources of Zn (98 +/- 13.8 ppm and 55 +/- 6.7 ppm). Leguminous green fodders (stylosanthus, lucerne, cow pea, soyabean) were excellent sources of Ca (1.9% +/- 0.16%), Mg (0.40% +/- 0.05%), Cu (30 +/- 5.2 ppm), Zn (121 +/- 14.7 ppm) and Fe (1234 +/- 166 ppm) and moderate sources of P. Mixed local grasses and weeds were high in silica (6.9% +/- 1.00%) but were good sources of Cu, Zn and Fe. Cereal grains (maize, wheat, rice, ragi) were low in ash (2.9% +/- 0.33%) and were relatively poor sources of Ca (0.22% +/- 0.03%), Mg (0.19% +/- 0.03%) and Cu (13 +/- 3.1 ppm). Pulses were low to medium sources of most minerals and good sources of Fe (1230 +/- 293 ppm). Oil seed cake/extractions (groundnut cake, cotton seed cake, soyabean meal, sunflower cake, safflower cake) and cereal by-products (rice polish, rice bran, wheat bran) were excellent sources of P (1.1% +/- 0.47% and 2.3% +/- 0.19%) and good sources of Zn (65 +/- 3.9 ppm and 66 +/- 10.7 ppm) and Fe (938 +/- 130 ppm and 662 +/- 126 ppm). Among the unconventional feeds screened, orange peel, sunflower heads, meat meal, rubber seed cake, spirulina algae and sea weeds contained plentiful Ca, Zn and Fe: tree leaves/top feeds (mulberry, erythrina, glyricidia, banana, subabul, groundnut haulms) were excellent sources of Ca (1.5% +/- 0.13%), Zn (120 +/- 22.9 ppm) and Fe (1033 +/- 133 ppm) but relatively poor sources of P. Soyabean husk, cocoa seed husk, rubber seed cake and meat meal were moderate to good sources of P (1.0% and 0.33%). The high Zn and Fe values of most feeds/fodders were probably due to soil contamination. This account of the micronutrient content of feed/fodder resources should help in strategic supplementation intended to alleviate local deficiencies. 相似文献
16.
Troxel MT Vite CH Van Winkle TJ Newton AL Tiches D Dayrell-Hart B Kapatkin AS Shofer FS Steinberg SA 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2003,17(6):850-859
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
17.
Bovine Mastitis in Selected Areas of Southern Ethiopia 总被引：3，自引：0，他引：3
A study on bovine mastitis, designed to determine the causal agents, prevalence of infection and impact of risk factors in three cattle breeds, was conducted in selected areas of southern Ethiopia. A total of 307 lactating and non-lactating cows, of which 162 were indigenous Zebu, 85 Jersey and 60 Holstein-Friesian, were examined by clinical examination and the California mastitis (CMT) test. Of these, 40.4% were positive by CMT and bacteriology for clinical or subclinical mastitis, with prevalence rates of 37.1% and 62.9%, respectively. Out of 1133 quarters examined, 212 (18.7%) were found to be infected, 83 (39.2%) clinically and 129 (60.8%) subclinically. The prevalence of mastitis was significantly higher in Holstein-Friesian than in indigenous Zebu, in non-lactating cows than in lactating cows, in the early lactation stage than in the mid-lactation stage, in cows with lesions and/or tick infestation on skin of udder and/or teats than in cows without this factor, and in the wet season than in the dry season. Mastitis increased with parity number (R = 0.9). Of 248 CMT and clinically positive udder quarter samples analysed microbiologically, 212 were culturally positive for known mastitis pathogens and 36 were negative. Of the 199 positive samples, Staphylococcus accounted for 39.2%, Streptococcus for 23.6%, coliforms for 14.1%, Micrococcus and Bacillus species for 8.0% each and Actinomyces or Arcanobacterium (Corynebacterium) for 7.0%. It was concluded that there was a high prevalence of clinical and subclinical mastitis, mainly caused by Staphylococcus aureus, Streptococcus agalactiae and Escherichia coli, in this study area. 相似文献
18.
不同日粮氮水平对山羊氮代谢和微生物蛋白质合成的影响 总被引：2，自引：0，他引：2
本试验旨在探讨不同日粮氮水平对内蒙古白绒山羊氮代谢和微生物蛋白质合成(MCP)的影响,以达到提高氮利用率,减少环境污染和饲料资源的浪费。试验选用9只体况良好、体重为(43.83±2.95)kg,装有瘤胃瘘管的内蒙古白绒山羊,按体重随机分为3组,每组3只。日粮分为低氮7.5%、中氮10.5%、高氮13.5%3个氮水平。试验使用常规试验方法分析测定粪尿血液等生化指标,通过全收尿法和尿嘌呤衍生物法估测MCP。结果表明:随着氮水平增加,氨氮(NH3-N)浓度、氮摄入量、尿氮排出、尿中尿素氮(UUN)和MCP显著增加(P0.05),干物质采食量(DMI)显著降低(P0.01);沉积氮占总摄入氮的比例在中氮组最高,MCP在高氮组最高。综上所述,适当降低日粮氮水平可减少粪尿氮排放,提高反刍动物氮利用率。 相似文献
19.
Emma Scurrell Sue Manning Pedro Malho Alex Civello John Mould Rossella Carrozza Bart E. Wagner 《Veterinary ophthalmology》2020,23(2):402-408
A microsporidial keratopathy is described in two dogs. Both dogs presented with a unilateral stromal keratopathy characterized by multifocal coalescing opacities, and the diagnosis was made on histopathologic examination of keratectomy specimens. Transmission electron microscopy (TEM) on formalin‐fixed, paraffin‐embedded corneal tissue was performed in one dog, and the morphologic features were consistent with Nosema species infection. Both dogs were initially diagnosed and treated by superficial keratectomy. One dog received additional antifungal medication and underwent a penetrating keratoplasty following local recurrence two years later. No other systemic lesions attributable to the microsporidial infection were identified clinically. The clinical and diagnostic pathology findings, treatment, and follow‐up are discussed. 相似文献
20.
优质肉鸡S3系体重与体尺性状指标的主成分分析 总被引：1，自引：0，他引：1
为研究地方优质肉鸡S3系体重和体尺性状之间的内在相关性,12周龄时选取同一饲养条件下的150只鸡(公母各半)进行体重和体尺性状等7个指标的测定。结果表明:S3系公鸡的体重、体斜长、龙骨长、胸角均显著高于母鸡(P0.05);公母鸡体重、体斜长、胸宽、胫围这4项指标变异系数较大,有较大选育空间;体重与体斜长、龙骨长、胸宽、胸角、胫围呈极显著正相关(P0.01),相关系数分别为0.57、0.52、0.51、0.49和0.47,与胸深、胫长呈显著相关(P0.05),相关系数分别为0.44和0.39;其他各体尺指标间也存在不同程度的正负相关性(P0.05或P0.01);体重和7个体尺性能指标综合成5个复合指标,累计贡献率达92.6355%,其中第l、2、3、4、5主成分分别解释总变异的36.5353%、22.8642%、13.3115%、10.4535%和8.4710%;各主成分的特征根分布较广,经统计计算入选的5个主成分所包含信息的侧重点各有相同,分别都在一定程度上反映了S3鸡的体型特征和生长发育规律,为今后选育出更加符合市场需求的屠宰加工优质型肉鸡指明了的方向。 相似文献