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101.
对原核表达的重组建鲤组织蛋白酶L(Cathepsin L,CAT L)蛋白进行尿素洗涤和Ni-NTA亲和层析纯化,该目的蛋白经300 mmol/L咪唑洗脱为单一峰,SDS-PAGE结合TSK-GEL G2000SWxl凝胶过滤高效液相色谱分析表明重组CAT L获得了高度纯化,分子量约28 k D,纯度超过95%。Z-Phe-Arg-MCA底物测活法显示该重组CAT L表现为半胱氨酸蛋白酶活性,能与其内源抑制因子Cystatin以1︰1的摩尔比结合,具有生物学活性。以纯化的重组CAT L蛋白免疫Balb/C小鼠获得抗血清,经ELISA法检测获得的CAT L抗血清效价高于1︰512000;Western blotting鉴定结果表明该抗体具有良好的特异性,能够识别原核表达的重组CAT L蛋白。免疫组织化学分析结果表明,该抗体还能识别建鲤小肠、肝胰脏、脾、背肌和心肌组织表达的内源性CAT L蛋白。因此可利用该抗体从蛋白水平检测CAT L在鱼类不同组织中的表达和分布情况。 相似文献
102.
Dong-Hui Zhou Fu-Rong Zhao Hui-Yan Xia Min-Jun Xu Si-Yang Huang Hui-Qun Song Xing-Quan Zhu 《Irish veterinary journal》2013,66(1):1-4
Chlamydia spp. are obligate intracellular gram-negative bacteria that cause a wide range of significant diseases in humans and animals worldwide, resulting in significant economic losses. Chlamydial infection in cattle has been reported in many countries including China. However, there has been no survey of chlamydial infection of dairy cattle in Guangzhou, southern China. The objective of the present investigation was to examine the chlamydial seroprevalence in dairy cattle in Guangzhou, subtropical southern China by using an indirect hemagglutination assay (IHA). The overall seroprevalence of chlamydial infection in dairy cattle was 7.25% (29/400). Greater than or equal to eight-yr-old dairy cattle had the highest seroprevalence (10.34%), followed by those that were ≥ 6 years old or < 7 years old dairy cattle (10.20%), although there were no statistically significant differences among different groups (P > 0.05). Dairy cattle with 5 pregnancies had the highest seroprevalence (10.81%). These results indicate that chlamydial infection was present in dairy cattle in Guangzhou, subtropical southern China, and integrated strategies and measures should be executed to control and prevent chlamydial infection and disease outbreak in the study region. 相似文献
103.
本试验通过研究免疫状况良好的母猪群所产仔猪获得母源抗体的方式,仔猪体内母源抗体的持续时间,仔猪不同免疫程序的免疫效果等,从而制定出仔猪猪瘟的最佳免疫程序。 相似文献
104.
Msoffe PL Minga UM Olsen JE Yongolo MG Juul-Madsen HR Gwakisa PS Mtambo MM 《Tropical animal health and production》2001,33(4):341-354
A study was conducted to determine the variations in physical characters and immunocompetence among scavenging local chicken ecotypes in Tanzania. Eighty-four adult scavenging local chickens from four eco-climatic regions of Tanzania were studied. Measurements of adult body weight, body length, shank length and egg weight and observations of plumage colour and pattern, earlobe colour, skin colour and the shape of the comb were conducted. The antibody response to sheep red blood cells, serum haemolytic complement and the cutaneous response to phytohaemagglutinin-P were assessed. Five ecotypes were identified and named Mbeya, Morogoro-medium, Ching'wekwe, Kuchi and Singamagazi. Singamagazi and Kuchi were significantly heavier, with longer shanks and heavier eggs than the other ecotypes. The average adult body weight for males ranged from 1621 g (Mbeya) to 2915 g (Singamagazi). Average female weights ranged from 1108 g (Morogoro-medium) to 2020 g (Singamagazi). Mean egg weights ranged from 37.65 g (Ching'wekwe) to 45.61 (Singamagazi). The Kuchi had mostly rose and walnut combs, while the other ecotypes were mostly single combed. In each ecotype there were chickens with a high or low antibody response to red blood cells, but there was a significant difference between the ecotypes. 相似文献
105.
近年来,从华东地区患腹泻仔猪中分离到一些表达K88菌毛的大肠杆菌,这些菌株只与K88a因子单抗反应,而不与b、c、d因子单抗反应。通过K88常规血清交叉吸收试验、SDS-PAGE、Western印迹,表明这些菌株不仅与K88ac参考菌株C83907制备的c因子血清反应,而且与以分离株SEC586制备且经K88ab、K88ac、K88ad参考菌株吸收后的血清也反应。对分离株SEC586、SEC464的K88主要亚单位结构基因faeG的克隆、测序,发现该基因由846对核苷酸组成,编码菌毛主要亚单位的262个氨基酸及21个氨基酸的信号肽,比国外报道的K88ac FaeG亚单位(263个氨基酸)少了1个氨基酸,比K88ab、K88ad(265个氨基酸)少了3个氨基酸。SEC586、SEC464菌株的FaeG亚单位氨基酸序列的同源性为97.7%,它们与K88ac的同源性为94.7%和96.2%;与K88ab的同源性为90.1%和91.2%;与K88ad的同源性为87.0%和88,6%。结果表明,新分离的K88ac大肠杆菌黏附素主要亚单位已发生了部分变异。 相似文献
106.
L. Daroda K. Hahn D. Pashkoulov E. Benvenuto 《Physiological and Molecular Plant Pathology》2001,59(6):317
The activation of Fusarium moniliforme endopolygalacturonase (endoPG) was studied during infection of maize plants. EndoPG is a plant cell wall degrading enzyme that cleaves the pectin component causing cell death. The authors generated several hybridoma cell lines producing endoPG specific monoclonal antibodies. One monoclonal antibody was selected and successfully used in Western blotting analysis to detect F. moniliforme endoPG secretion in vitro and in planta. Two F. moniliforme strains (FC-l0 and 62264) were used for the studies. Both strains revealed the expression of a single endoPG in vitro as in planta. EndoPG from strain FC-10 presented four isoforms whereas only two isoforms were revealed in the endoPG from strain 62264. Differences were also found in the sequences of the two endoPG genes indicating the presence of endoPG variability among F. moniliforme strains. 相似文献
107.
Kenichi?TsuchiyaEmail author Christophe C.M.?D’Ursel Mitsuo?Horita Yuzo?Nozu 《Journal of General Plant Pathology》2003,69(5):310-315
Strains of Xanthomonas campestris pv. vesicatoria Dye 1978 (Xcv), the causal agent of bacterial spot, have been classified into two groups based on their ability to hydrolyze starch. Three monoclonal antibodies (MAbs), 7AH10, 5HB3, and 4AD2, were produced immunized against the living bacteria and were specific to and could distinguish Xcv strains able or unable to hydrolyze starch (Amy+ or Amy–). The MAb 7AH10, obtained against strain UPB141(Amy–) reacted in an enzyme-linked immunosorbent assay with all the Amy– strains (n = 19) and 1 of 11 Amy+ strains. Against Xcv 2625, an Amy– unusual phenotype strain, MAb 5HB3, recognized 97% of our worldwide collection of Xcvs (n = 30). Also against that strain, the MAb 4AD2 reacted with none of the homologous Amy– phenotypes and with 90% (n = 11) of the heterologous Amy+ phenotypes. For all the MAbs, cross reactions with other pathovars or species were less than 4% (n = 67). By assaying a Japanese collection of strains against the three MAbs, the Amy+ strains were distinguished from the Amy– strains, and their relation with other world strains could be demonstrated. All the MAbs reacted with the lipopolysaccharide fraction of the bacterial cell wall during immunoblotting. 相似文献
108.
Atkinson RA Cook RW Reddacliff LA Rothwell J Broady KW Harper P Ellis JT 《Australian veterinary journal》2000,78(4):262-266
OBJECTIVE: To investigate the seroprevalence of Neospora caninum infection in a commercial dairy cattle herd, 15 months after detection of an abortion outbreak. PROCEDURE: Sera from the whole herd (n = 266) were examined for N caninum antibodies by indirect fluorescent antibody test (IFAT) and immunoblot analysis. Herd records were reviewed to collate serological results with abortion history, proximity to calving, and pedigree data. RESULTS: The seroprevalence of N caninum infection was 24% (63/266) for IFAT titre > or = 160, 29% (78/266) for immunoblot positive (+ve), and 31% (82/266) for IFAT > or = 160 and/or immunoblot +ve; 94% (59/63) of animals with IFAT > or = 160 were immunoblot +ve. The association between seropositivity (IFAT > or = 160 and/or immunoblot +ve) and history of abortion was highly significant (P < 0.001); the seroprevalence was 86% (18/21) in aborting cows, compared with 30% (50/164) in non-aborting animals. The abortion rate for seropositive cows was 26% (18/68) compared with 3% (3/117) for seronegative animals. IFAT titres of infected cows were higher within 2 months of calving than at other times (P < 0.001). The association between seropositivity in dams and daughters was highly significant (P = 0.009). CONCLUSIONS: The abortions were associated with N caninum infection and there was evidence of reactivation of latent infection close to calving and congenital transmission of infection. Immunodominant antigens identified by immunoblots may prove useful for improved diagnostic tests. 相似文献
109.
Tropical Animal Health and Production - 相似文献
110.
Kruger JM Venta PJ Swenson CL Syring R Gibbons-Burgener SN Richter M Maes RK 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2000,14(6):593-597
The gammaherpesvirus bovine herpesvirus-4 (BHV-4) has been isolated from a wide variety of animals, including lions and domestic cats. Although BHV-4 antibodies have been detected in normal cats and cats with urinary disorders, the epidemiology and pathogenic role of BHV-4 in cats is unknown. The purpose of this study was to determine the prevalence of BHV-4 antibodies and viral nucleic acid in a population of free-roaming cats. Plasma and peripheral blood leukocyte samples were collected from 52 male and 52 female free-roaming cats impounded at a regional animal control facility in Central Michigan. Plasma concentrations of BHV-4 antibodies were measured with an indirect fluorescent antibody test. Peripheral blood leukocyte DNA was isolated, and a 2-stage polymerase chain reaction with heminested primers delineating a conserved portion of the BHV-4 glycoprotein B gene homologue was used to amplify BHV-4-specific DNA sequences. BHV-4 antibodies were detected in 38 (73%) male and 23 (44%) female cats. Seropositive cats were significantly more likely to be male than female (odds ratio = 3.22; P = .007). Cell-associated viremia was detected in 17 (33%) male and 11 (21%) female cats. Of the 61 seropositive cats, 23 (38%) had a detectable viremia; only 5 (12%) seronegative cats had detectable viremia. Seropositive cats were significantly more likely to be viremic than seronegative cats (OR = 4.30: P = .009). Our results suggest that BHV-4 infection may be more widespread in certain cat populations than previously reported. Furthermore, many cats seropositive for BHV-4 antibodies have a concurrent cell-associated viremia. 相似文献