共查询到20条相似文献，搜索用时 500 毫秒
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金黄色葡萄球菌是引起奶牛乳腺炎的主要传染性病原菌.本实验采用分子克隆及蛋白表达技术,成功的表达了金黄色葡萄球菌纤维素结合蛋白A(FnBPA)D片段并对其免疫学活性进行了初步研究.表达产物免疫实验动物后,获得较高效价的抗体.用制备血清进行吞噬调理试验,抗金黄色葡萄球菌黏附等一系列试验,证明 抗体对金黄色葡萄球菌具有吞噬调理作用,也有抗金黄色葡萄球菌黏附的作用,这将为制备奶牛乳房炎金黄色葡萄球菌黏附素疫苗奠定基础. 相似文献
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纤维素结合蛋白B(FnbpB)是金黄色葡萄球菌(S.aureus)最主要的黏附因子之一,它能够介导S.aureus结合于细胞表面的纤维连结素(Fn)和纤维蛋白原(Fg)。FnbpB基因中D区为主要活性部位。本实验利用pET-32a表达载体表达了S.aureus FnbpB-D重组蛋白(34.7ku),并通过动物实验对重组FnbpB-D蛋白的抗血清活性进行了初步研究。采用ELISA对抗血清进行抗粘附性检测,结果显示实验组与对照组差异极显著(p0.01),抗血清具有较强的抑制S.aureus黏附纤维连结素的作用。此外,调理吞噬实验结果显示,免疫兔全血对S.aureus有较强的调理作用。这些实验结果将为制备奶牛乳房炎S.aureus黏附素疫苗的研制提供参考。 相似文献
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<正> 一、前言调理素 B(Ⅱ)是从药用植物中提取的一种非激素性的细胞激动剂,它能疏通微循环,促进细胞新陈代谢。据称在饲料中添加调理素,可改变其适口性,提高饲料利用率。为验证这一说法,我们于85年12月至86年1月在江西农大教学实验站进行了 相似文献
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猪链球菌-副猪嗜血杆菌二联亚单位疫苗在小鼠的免疫效力分析 总被引：1，自引：0，他引：1
本研究旨在开发一种用于防制猪链球菌病和副猪嗜血杆菌病的新型二联基因工程疫苗,并对其免疫效应进行评价.选择猪链球菌2型蛋白溶血素SLY、HP0197和副猪嗜血杆菌外膜蛋白D15和HPS06257 4种蛋白,重组表达并纯化,添加一定比例的佐剂制成含有不同组分的亚单位疫苗.研究表明该二联亚单位疫苗及各单苗能够引起小鼠良好的体液免疫,并能保护机体分别抵抗致死剂量猪链球菌2型(SS2)和副猪嗜血杆菌(HPS)强毒株的攻击.此外,调理吞噬试验表明疫苗高免血清能够有效杀死细菌,并赋予机体被动抵抗SS2和HPS感染的能力.研究证实制备的猪链球菌-副猪嗜血杆菌二联亚单位疫苗在小鼠具备良好的免疫保护效力. 相似文献
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利用碱性蛋白酶水解家蚕丝素蛋白制备抗氧化肽,考察水解pH、水解时间、加酶量、底物浓度、水解温度5个因素对水解产物DPPH自由基清除率的影响,在此基础上采用响应面试验优化水解工艺条件,并调查家蚕丝素蛋白抗氧化肽对热、酸、碱和体外肠道消化的稳定性,测定不同分子质量范围的家蚕丝素蛋白水解产物组分的抗氧化活性.结果 表明,碱性蛋白酶水解制备家蚕丝素蛋白抗氧化肽的最优工艺条件为水解pH 8.69、水解时间60 min、加酶量3035 U/g、底物(丝素蛋白)质量浓度26.80 g/L、水解温度53.89℃,在此条件下制备获得的家蚕丝素蛋白抗氧化肽对DPPH自由基的清除率为55.16％.家蚕丝素蛋白抗氧化肽具有良好的热稳定性;在中性环境中抗氧化活性很稳定,但在强酸和强碱环境中抗氧化活性明显降低;经体外模拟胃肠道消化后,对DPPH自由基的清除率比未消化处理对照组提高了9.43百分点.分子质量小于5 kD组分是家蚕丝素蛋白抗氧化肽的主要活性组分.研究结果为家蚕丝素蛋白功能产品的开发提供了试验依据. 相似文献
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猪链球菌2型的致病特性与毒力因子 总被引：27，自引：3，他引：24
介绍了猪链球菌２型对猪人、人、其他动物及实验动物的致病特性;概述了猪链球菌溶菌酶释放蛋白（ＭＲＰ）、细胞外蛋白因子（ＥＦ）、溶血素、英膜多糖、４４ｋｂ蛋白和ＩｇＧ结合蛋白及纤毛、粘着素等毒力因子的研究概况。 相似文献
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随着动物性蛋白的日趋减少,人们越来越重视植物性蛋白营养利用的限制因素。本文着重研究生长家畜在消化、吸收和利用植物性蛋白过程中的一些问题。消化植物性蛋白不同于动物性蛋白,它不易分解,经加热处理消化率虽所有提高,但大部分仍不易消化。植物性饲料中的某些物质对肠道消化酶具有直接的抑制作用,降低消化速度。但对蛋白酶抑制素的确切作用目前尚不清楚。众所周知,大豆中至少有三种主要的蛋白酶抑制素:经典的,即 Kunitz 抑制素、高胱氨酸Bowman-Birk 抑制素和一些低分子量抑制素.此外,大豆中还有不同数量的酚和鞣酸, 相似文献
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A bovine serum protein, initially recognized by its inhibitory effect on the hemolytic activity of the bovine alternative pathway was isolated from fresh bovine serum by polyethylene glycol precipitation and chromatography on DEAE-Sephacel, CM-Sephadex A-50 and Sephadex G-200. The protein, a single chain polypeptide with an apparent molecular weight of 158,000, was identified as factor H, a regulatory protein of the alternative complement pathway. Functional characterization of this protein as factor H was based on the following properties: binding to C3b, inhibition of factor B binding to C3b, cofactor activity in the cleavage of C3b by factor I, inhibition of fluid phase alternative pathway C3 convertase (C3b.Bb) formation and activity, and species-specific inhibition of the alternative pathway mediated hemolysis of heterologous erythrocytes. A monospecific rabbit antiserum against bovine factor H failed to react with human serum factor H. 相似文献
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C3 was obtained from bovine serum by polyethylene glycol precipitation and chromatography on DEAE-Sephadex A-50, CM-Sephadex A-50 and Sephacryl S-200. The protein has a molecular weight of 183,000 (alpha-chain 114,000 and beta-chain 69,000). A CVF-induced bovine C3 convertase (Sepharose-CVF.Bb) cleaved C3 into C3a (11,000) and C3b (172,000) as shown by SDS-polyacrylamide gel electrophoresis. Isoelectricfocusing of C3 demonstrated at least three electrophoretic variants with pI 6.55–6.85. The isolated protein promoted the formation and action of a C3 convertase in the presence of purified bovine factors B and D. A monospecific antiserum prepared in rabbits failed to cross react with human C3 or CVF. C3c was identified as a contaminant during the isolation of C3. 相似文献
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Spectrophotometry and Ultracentrifugation for Measurement of Plasma Lipids in Dogs with Diabetes Mellitus 
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下载免费PDF全文 E.C. Seage K.J. Drobatz R.S. Hess 《Journal of veterinary internal medicine / American College of Veterinary Internal Medicine》2018,32(1):93-98
Background
There are conflicting reports of plasma lipoprotein lipid content in dogs with diabetes mellitus (DM).Objectives
To determine lipoprotein lipid content of plasma of dogs with DM by spectrophotometry and ultracentrifugation; to compare lipoprotein lipid content in diabetic and healthy dogs; and to quantify apolipoprotein B‐100 (ApoB) in dogs with DM.Animals
22 dogs with DM and 9 healthy dogs.Methods
Cross‐sectional study. Triglyceride (TG), total cholesterol (TC), and high‐density lipoprotein cholesterol (HDL‐C) concentrations were measured by spectrophotometry. Very low‐density lipoprotein cholesterol (VLDL‐C) and low‐density lipoprotein cholesterol (LDL‐C) concentrations were calculated after ultracentrifugation. Non‐HDL‐C cholesterol was calculated by subtracting HDL‐C from TC. ApoB was quantified by ELISA. The Mann‐Whitney test was used for comparison of median lipoprotein concentrations, and Spearman's correlation was used to assess associations between ApoB and lipoprotein fractions.Results
All values are reported in mg/dL. Median TG (122), TC (343.5), HDL‐C, (200), VLDL‐C, (27) LDL‐C (68), non‐HDL‐C (114), and ApoB (320) were significantly higher in dogs with DM, compared to healthy dogs (57, 197, 168, 12, 16, 31, and 258, respectively, P‐values 0.0079, <0.001, 0.029, 0.011, <0.001, <0.001, 0.025, respectively). A significant association was found between ApoB and LDL‐C (Spearman's rho = 0.41, P = 0.022) and between ApoB and non‐HDL‐C (Spearman's rho = 0.40, P = 0.027).Conclusions and Clinical Importance
Dyslipidemia of dogs with DM is characterized by pronounced increases in LDL‐C and non‐HDL‐C concentrations, although all lipoprotein fractions are significantly increased. Knowledge of specific lipoprotein fraction alterations in dogs with DM can enhance treatment options for diabetic dyslipidemia in dogs. 相似文献15.
Takahashi Y Sato K Itoh F Miyamoto T Oohashi T Katoh N 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2003,65(2):199-205
Apolipoprotein E (apoE) is a protein constituent of lipoproteins, and acts as a receptor-binding ligand. Although the existence of bovine apoE in lipoprotein fractions has already been reported, quantitative studies on the changes of apoE in plasma and lipoprotein fractions are lacking. In the present study, an increase of a 38 kDa protein in the very low-density lipoprotein (VLDL) fraction obtained from fasted calves was detected. This 38 kDa protein was identified as bovine apoE by determination of the N-terminal amino acid sequence. Bovine apoE was purified and an enzyme-linked immunosorbent assay (ELISA) was developed. Using this system, the effect of fasting on the concentration of apoE in plasma and the distribution of apoE in lipoprotein fractions were investigated. After 3 days of fasting, the concentration of plasma apoE increased significantly (p<0.05) by 280 %, and was returned to the basal level by 3 days of refeeding. The lipoprotein fractions obtained from before and after fasting was separated by ultracentrifugation. ApoE was significantly increased in VLDL, low-density lipoprotein (LDL) and non-lipoprotein fractions by fasting (p<0.05). On the other hand, in high-density lipoprotein (HDL) fractions obtained from both before and after fasting, the level of apoE was very low compared to the other fractions. These results suggested that bovine apoE contents in triglyceride-rich lipoproteins are modulated by nutritional treatment and closely associated with triglyceride-rich lipoprotein metabolism. 相似文献
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家蚕30 kD脂蛋白家族在家蚕的生长发育过程中具有重要的生理功能。从家蚕蛹cDNA文库中克隆到一条编码30kD蛋白质的基因cDNA序列,该序列全长2 803 bp,ORF为792 bp,编码含263个氨基酸残基的脂蛋白(low molecular 30K lipo-protein pBmHPC-21;GenBank登录号:Q00801),命名为BmLp-c21。将BmLp-c21克隆至原核表达载体pET-28a(+),转化E.coli Rosetta(DE3),IPTG诱导表达后通过亲和层析得到纯化的融合蛋白,并免疫新西兰大白兔制备多克隆抗体。亚细胞定位显示BmLp-c21主要存在于细胞质中,呈点状或者片状分布。通过实时荧光定量PCR和Western blotting方法,分别检测到家蚕不同发育时期和5龄幼虫不同组织中BmLp-c21 mRNA的转录水平与蛋白质的表达水平存在较大差异,在蛹期及5龄幼虫血淋巴中的mRNA转录水平和蛋白质表达水平最高。初步推测BmLp-c21在家蚕蛹的变态发育过程以及蚕体脂质的运输方面具有重要的功能。 相似文献
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本试验旨在研究饲粮中添加不同水平苜草素对蛋鸡胆固醇代谢的影响,并探讨其基因表达调控机制。选取体重和产蛋率相近的26周龄尼克蛋鸡540只,随机分为5组,每组6个重复,每个重复18只。对照组饲喂基础饲粮,试验组分别在基础饲粮中添加300、600、900、1 200 mg/kg的苜草素。预试期7 d,正试期70 d。结果表明:1)试验第35天和第70天,900 mg/kg苜草素组的蛋黄胆固醇含量显著低于对照组(P0.05)。2)与对照组相比,900 mg/kg苜草素组显著升高了蛋鸡血清中高密度脂蛋白胆固醇(HDL-C)含量和高密度脂蛋白/低密度脂蛋白(HDL/LDL)(P0.05),各组蛋鸡血清中总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL-C)含量差异均不显著(P0.05)。3)与对照组相比,600、900、1 200 mg/kg苜草素组显著降低了蛋鸡肝脏中3-羟基-3-甲基戊二酸单酰辅酶A还原酶(HMGCR)mRNA表达量(P0.05),900和1 200 mg/kg苜草素组显著升高了蛋鸡肝脏组织中胆固醇7α-羟化酶(CYP7A1)mRNA表达量(P0.05)。各组蛋鸡肝脏中固醇结合蛋白元件-2(SERBP-2)和卵巢中卵黄蛋白原受体(OVR)mRNA表达量无显著差异(P0.05)。综上,饲粮中添加苜草素降低了蛋黄胆固醇和全蛋胆固醇含量,其调控机制可能是通过抑制蛋鸡胆固醇的内源性合成和促进胆固醇向胆汁酸的转化排泄2种途径来实现。本试验推荐26~35周龄蛋鸡苜草素添加水平为900 mg/kg。 相似文献
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Katoh N Oikawa S Oohashi T Takahashi Y Itoh F 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2002,64(1):51-55
Reduced feed intake near parturition is suggested to be one of the major causal factors for the development of fatty liver in cows, and nonfeeding has been used as an experimental model for fatty liver. In cows with fatty liver, concentrations of lipoprotein lipids and proteins are decreased. In addition, the acute-phase protein haptoglobin is induced. The purpose of the present study was to examine whether the decrease of lipoprotein concentrations and the induction of acute-phase proteins were similarly reproduced by non-feeding. Holstein female calves (n=5) were nonfed for 3 days and thereafter refed. Serum concentrations of nonesterified fatty acids and beta-hydroxybutyric acid were initially increased by the nonfeeding, and followed by decreases in concentrations of cholesteryl esters, phospholipids, apolipoprotein (apo) B-100 and apoA-I. The apoC-III concentration was not distinctly decreased. Haptoglobin and serum amyloid A were induced during the nonfeeding and refeeding process. Haptoglobin was distributed in different proportions in the high-density lipoprotein, very high-density lipoprotein and the lipoprotein-deficient fractions, whereas almost all serum amyloid A was associated with the high-density lipoprotein fraction. These results suggest that the decreases in lipoprotein concentrations and induction of acute-phase proteins found in cows with fatty liver and those with fatty liver-related diseases such as ketosis are primarily due to the reduced feed intake near parturition. 相似文献
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The agarose gel electrophoresis described by Johansson (1972) was modified so that a buffer of pH 7.9 was used in the gel, whereas the buffer in the electrode vessels had a pH of 8.6.The cattle blood serum protein picture is described in detail. The β1-globulin zone shows a very distinct picture of the genetically polymorphic bovine transferrins. The region between the α- and β-globulins shows a number of faint and often very distinct bands. A faint background staining over the whole electrophoretogram may partly be caused by a rather strong lipoprotein in the α1-region, lipids thus having migrated all over the electrophoretogram.The modified method described is well suited as a “screen electrophoresis” for cattle serum and is also useful e.g. in studying bovine transferrin polymorphism. 相似文献
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Thyroxine-binding prealbumin (TBPA) and thyroxine-binding globulin (TBG) were isolated from canine serum and partially characterized. TBPA was isolated by retinol-binding protein (RBP) affinity chromatography and further purified by preparative agarose gel electrophoresis or FPLC ion exchange chromatography. TBG was purified by thyroxine (T4)-Sepharose chromatography followed by gel filtration on Sephacryl S-300 and preparative electrofocusing in a granulated dextran gel. Molecular weights were estimated by SDS-polyacrylamide gradient gel electrophoresis. Canine TBPA had a tetramer molecular weight of 56,000, an extinction coefficient of 12.8 cm2cg−1, an isoelectric point of 5.26–5.70 and a microheterogeneity pattern similar to that of human TBPA. Partial immunochemical identity with human TBPA was also found. Plasma concentrations of TBPA were measured by rocket immunoelectrophoresis in 43 normal and 35 hypothyroid dogs. Reference levels for TBPA ranged between 205 and 474 mg/l. Hypothyroid dogs had a mean TBPA level of 315.0 mg/l (SD: 91.1 mg/l). TBG had a molecular weight of 75,000 and an isoelectric point of 5.0. No immunochemical identity with human TBG was found. Gel filtration of serum on Sephacryl S-200, identification of T4-binding proteins with 125I-T4, and protein- and lipoprotein staining of fractions was performed. Thyroxine-binding was found to TBG in the β-globulin region, TBPA in the 2-region, albumin, and to the high density lipoprotein (HDL2) in the 1-region and the very low density lipoprotein (VLDL) in the pre-β region. A corresponding band to the latter protein in serum was masked by TBG and TBPA, and T4-binding in the 1-region was not always seen in serum. Many similarities were found between man and dog regarding TBPA, but not TBG. The differences in structure of TBG may in part be responsible for the low serum T4 levels and rapid T4 metabolism seen in dogs. 相似文献