旋毛虫肌幼虫可溶性抗原、排泄分泌抗原的电泳分析

本文报道了应用十二烷基硫酸钠一聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ），聚丙烯酰胺凝胶等电聚焦（ＩＥＦ）电泳及二维电泳（ＩＥＦ／ＳＤＳ－ＰＡＧＥ）对旋毛虫肌动虫排泄分泌（ＥＳ）抗原和肌幼虫可溶性抗原的分析结果。肌幼虫ＥＳ抗原经ＳＤＳ－ＰＡＧＥ后用考马斯亮蓝染色蛋白质，结果显示１６条蛋白带，分子量范围２１～８０ＫＤ，其中主带９条。ＩＥＦ电泳后分别用ＰＡＳ染多糖、考马斯亮蓝Ｒ－２５０染蛋白质、Ｎｉｌｅ＇ｓ蓝染脂、醋酸ａ－萘酯／坚固蓝染酪酶同工酶，结果肌幼虫ＥＳ抗原分别显示１６，２６、７及０条带；肌幼虫可溶性抗原分别显示２１、３８、４及１１条带。二维电泳后用考马斯亮蓝Ｇ－２５０染色多肤斑点，结果ＥＳ抗原显示多肽斑点６１个；肌幼虫可溶抗原显示１２２个多肽斑点。     

旋毛虫肌幼虫可溶性抗原,排汇分泌抗原的电泳分析

枉文报道了应用十二烷基硫酸钠－聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ），聚丙烯酰胺凝胶等电聚焦（ＩＥＦ）电泳及二维电泳（ＩＥＦ／ＳＤＳ－ＰＡＧＥ）对旋毛虫肌幼虫排汇分泌（ＥＳ）抗原和肌幼虫可溶性抗原的分析结果。肌幼虫ＥＳ抗原经ＳＤＳ－ＰＡＧＥ后用考马斯亮蓝染色蛋白质，结果显示１６条蛋白带，分子量范围２１￣８０ＫＤ，其中主带９条。ＩＥＦ电泳后分别用ＰＡＳ染多糖、考马斯亮蓝Ｒ－２５０染蛋白质、Ｎｉｌｅ＇     

牦牛肝片吸虫分泌排泄抗原的生化特性和免疫印迹分析

肝片吸虫的分泌排泄抗原（ＥＳ抗原）在诊断及诱导机体产生抗体方面具有重要作用。本文对寄生于牦牛的肝片吸虫ＥＳ抗原的蛋白质组成及其生化特性进行了分析。ＳＤＳ聚丙烯酰胺凝胶电泳显示ＥＳ抗原共有９条带，主要由１２－２６ＫＤ的小分子量蛋白质组成；糖蛋白及脂蛋白染色后发现ＥＳ抗原中糖蛋白极少，而含有较多的脂蛋白；等电聚焦电泳结果显示ＥＳ抗原有２２条带，几乎全部是酸性蛋白，ｐＩ主要集中在４．２５～５．２５范围内；双向电泳共检出３０个多肽，主要分布在ｐＩ４．５～７．０之间；免疫印迹分析结果表明：ＥＳ抗原中分子量为１６．２ＫＤ～１８．６ＫＤ的蛋白质是主要的抗原成分，其中１７．２ＫＤ多肽具有最强的免疫原性。     

肝片吸虫成虫抗原蛋白组分的研究

应用十二烷基磺酸钠一聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）电泳转移和固相免疫酶测定相结合的酶联免疫印溃技术（ＥＴＩＢ）首次分析了牛肝片吸虫成虫抗原白组分，结果显示：牛肝片吸虫成虫抗原分离出４４条带，经ＥＴＩＢ分析，抗原与阳性血清反应显示１８条带，其中主带有４条，分子量分别为３５、８０、９２和９４ＫＤ。上述蛋白组分有较强的反应性，可用作肝片吸虫的诊断抗原。     

用大片吸虫（Fasciola gigantica，Fg）成虫的28－kDa半胱肽水解酶进行反刍动物体内片吸虫病的免疫诊断（摘要）

用大片吸虫（Ｆａｓｃｉｏｌａｇｉｇａｎｔｉｃａ，Ｆｇ）成虫的２８－ｋＤａ半胱肽水解酶进行反刍动物体内片吸虫病的免疫诊断（摘要）Ｆａｇｂｅｍｉ，Ｂ．Ｏ．＆Ｇｕｏｂａｄｉａ，Ｅ．Ｅ．本文介绍一种以Ｆｇ成虫的２８ｋＤａ半胱肽水解酶作为片吸虫的特异性抗原，建...     

半胱胺对小鹅血浆中β-END和某些激素的影响

选用２４只二月龄的杂交鹅（川白×太湖）,随机分成对照组（１２只）和试验组（１２只）。试验组于日粮中一次性添加半胱胺（１００ｍｇ／ｋｇ·ＢＷ）。处理后第三天从翅静脉采取血样。用ＲＩＡ双抗法测定其中的β－ＥＮＤ、ＩＧＦ－Ｉ和各种激素的含量。结果表明：试验组的ＳＳ含量较对照组低１９．６４％（Ｐ＜０．０５）、而ＧＨ、ＩＧＦ－Ｉ和β－ＥＮＤ含量较对照组分别高５０．００％（Ｐ＜０．０１）、６２．８０％（Ｐ＜０．０１）和４４．５５（Ｐ＜０．０５）;与对照组相比较,试验组的ＴＳＨ低３２．３２％（Ｐ＜０．０５）,Ｔ４下降１４．７２％（Ｐ＞０．０５）,而Ｔ３升高２６．８０％（Ｐ＜０．０１）。以上结果提示：半胱胺可能是通过降低鹅血液中ＳＳ含量,使β－ＥＮＤ、ＩＧＦ－Ｉ、ＧＨ和Ｔ３水平升高,从而促进了鹅的快速生长。     

水牛梭形住肉孢子虫包囊纯化抗原的SDS—PAGE分析

应用ＳＤＳ－ＰＡＧＥ分析了水牛梭形住肉孢子虫包囊纯化抗原的蛋白质组分。采用垂直板型电泳，连续凝胶系统法，以考马斯亮兰Ｒ－２５０染色进行ＳＤＳ－ＰＡＧＥ分析的结果表明，水牛梭形住肉孢子虫包囊纯化抗原至少由１０种蛋白质组成，分子量范围为１７．５ＫＤ～１３５ＫＤ。其中主要蛋白质组分有５种，分子量分别为１２５ＫＤ、９８ＫＤ、９０ＫＤ、６９ＫＤ及３５ＫＤ。     

ABC—Dot—ELISA检测鸡传染性支气管炎病毒

建立了ＡＢＣ－Ｄｏｔ－ＥＬＩＳＡ检测鸡传染性支气管炎病毒（ＩＢＶ）的方法,其最佳反应条件是：包被液为０．０５ｍｏｌ／ＬｐＨ９．６的碳酸钠－碳酸氢钠缓冲液（ＣＢＳ）,免疫ＩＢＶ ＩｇＧ的最佳工作浓度为１：８０;抗原的最佳浓度为１：８００;封闭剂选用０．０１ｍｏｌ／Ｌ ｐＨ７．４含３０ｍＬ／Ｌ白明胶的ＰＢＳ,Ｂ－ＡｇＧ和ＡＢＣ－ＨＥＰ的最佳工作浓度为１：２００。用ＡＢＣ－ＨＥＰ的最佳工作浓度为１：２０     

抗鸡IgG重链单克隆抗体的制备与应用

以提纯鸡ＩｇＧ做抗原免疫Ｂａｌｌ／ｃ小鼠，取鼠细胞在ＰＥＧ１０００作用下与小鼠骨髓细胞（Ｓｐ２／ｏＡｇ１４）融合，采用间接免疫荧光（ＩＦＡ）和酶联免疫吸附试验（ＥＬＩＳＡ）检测上清抗体，阳性孔经有限稀法进行细胞克隆，共获得了７株分泌抗鸡ＩｇＧ单克隆抗体的杂交瘤细胞（２Ｈ８、４Ｄ８、４Ｄ８、１Ｂ７、２Ａ７、２Ｂ３、１Ｇ１２、３Ｄ１２）将这些细胞分别接种间系小鼠制备出腹水抗体，ＥＬＩＳＡ效价可达１０＾     

抗大片吸虫（Fg）的28－kDa水解酶单抗的建立及鉴定（摘要）

抗大片吸虫（Ｆｇ）的２８－ｋＤａ水解酶单抗的建立及鉴定（摘要）Ｆａｇｂｅｍｉ，Ｂ．Ｏ．用从大片吸虫Ｆｇ中提取的部分纯和完全纯的２８－ｋＤａ水解酶，高免ＢＡＬＢ／Ｃ小鼠，得到抗Ｆｇ的２８－ｋＤａ水解酶的单抗，我们利用ＦＡＳＴ－ＥＬＩＳＡ试验，免疫斑点试...     

多头绦虫抗原特性的研究──多头绦虫节片抗原及原头节ES抗原的SDS-PAGE分析

为了探明多头绦虫成虫节片抗原及多头蚴原头节排泄分泌（ES）抗原的多肽组分，以供今后在免疫诊断与预防脑多头蚴病的应用，本试验应用SDS-PAGE首次分析了多头绦虫成虫节片抗原及多头蚴原头节ES抗原的多肽组分。应用12．5％凝胶的连续系统，垂直板型电泳，考马斯亮兰R－250染色的结果表明，成虫节片抗原共有16条多肽带，其分子量范围29～154kD，其中主带4条，分别为73、88、128、134kD，原头节ES抗原共6条多肽带，分子量范围33～132kD，其中主带2条，分别为33和108kD。本试验初步阐明多头绦虫成虫节片抗原和原头节ES抗原的多肽组分，为进一步研究多头绦虫抗原的特性奠定了基础。     

兔狲血液蛋白质和酶的电泳分析

采用聚丙烯酰胺凝胶电泳法对２只兔狲的４种血液蛋白质表型和３种血液同工酶酶谱进行研究。结果发现：①被检兔狲的血红蛋白（ＨＢ），白蛋白（ＡＬＢ）和后白蛋白（Ｐａ）分别呈单一的ＨＢＡＢ，ＡＬＢＡＡ和ＰａＡＡ型，运铁蛋白（ＴＦ）有ＴＦＡＢ和ＴＦＢＢ两种表型；②血清碱性磷酸酶（ＡＬＰ）由ＡＬＰＡ，ＡＬＰＢ，ＡＬＰＣ和ＡＬＰＤ四种同工酶组成；③血清酯酶（ＥＳ）由ＥＳＩ和ＥＳＩＶ两种同工酶共７条区带组成；④血清乳酸脱氢酶（Ｓ－ＬＤＨ）由ＬＤＨ１，ＬＤＨ２，ＬＤＨ３，ＬＤＨ４和ＬＤＨ５五种同工酶组成，红细胞内只有前四种ＬＤＨ同工酶。     

Antigenic identification of excretory-secretory products of adult Dirofilaria immitis

Excretory-secretory (ES) products collected from adult Dirofilaria immitis cultured in vitro were analyzed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunoblotting. ES products of male (M-ES) and female (F-ES) worms were separated into 16 and 21 bands by SDS-PAGE with Coomassie blue and silver staining, respectively. The antigenic bands were then analyzed by immunoblotting, using pooled sera from dogs that had naturally contracted D. immitis. Sera from dogs with microfilaremic infection showed 7 bands in M-ES and 10 bands in F-ES, while those from dogs with occult infection revealed 3 bands in M-ES and 10 bands in F-ES. Among these bands, those of 14, 18, 21, 22, 29, and 32 kilodaltons (Kd) were common to M-ES and F-ES, those of 39 and 44 Kd were specific to M-ES, and those of 20, 38, 43, 53, 63, 90, 110, 125 and 136 Kd were specific to F-ES.     

高效毛细管电泳法同时检测饲料中七种防腐剂

建立酸性条件下同时分离测定山梨酸(SA)、苯甲酸(BA)、脱氢乙酸(DHA)、对羟基苯甲酸甲酯(MP)、对羟基苯甲酸乙酯(EP)、对羟基苯甲酸丙酯(PP)、对羟基苯甲酸丁酯(BP)七种防腐剂的高效毛细管电泳法。本试验用甲醇:水=1:1(体积比)的混合液作为提取剂提取饲料样品中防腐剂;以40 mmol/l磷酸二氢钾(KH2PO4)和100 mmol/l十二烷基硫酸钠(SDS)的1:1体积比混合液(pH=4.00)作为缓冲液,采用高效毛细管胶束电动色谱法测定样品中七种防腐剂。该方法在19 min内实现了七种防腐剂的分离;SA、BA的线性范围分别为1～500μg/ml和3～500μg/ml,DHA、MP、PP、BP、PP的线性范围均为5～500μg/ml,线性相关系数≥0.999 5。SA、BA、DHA和四种对羟基苯甲酸酯类防腐剂的检测限分别为0.5、1.5、3.0、2.5μg/ml;样品平均回收率为80.8%～107.0%,相对标准偏差≤5.0%。该方法高效快速分离了多种防腐剂,并且可以应用到各种饲料样品的检测。     

Effects of a unique application of electrical stimulation on tenderness, color, and quality attributes of the beef longissimus muscle

This study evaluated effects of four uniquely applied beef carcass electrical stimulation (ES) treatments on USDA grade factors, muscle color, subprimal purge loss, cooked steak weight loss, and cooked steak tenderness. One side of each (n = 284) beef carcass was subjected to ES using one of four treatments (medium voltage for medium duration, MVMD; medium voltage for long duration, MVLD; high voltage for medium duration, HVMD; or high voltage for long duration, HVLD) and was compared to its corresponding non-ES control side. Electrical stimulation of beef sides was applied focusing on middle meats while preventing severe contraction of the round and chuck. From matched (ES and control) sides of 120 carcasses (10 each of Select, low Choice, and upper two-thirds of Choice in each of the four ES treatments), longissimus steaks (2.5 cm thick) were cooked and used for Warner-Bratzler shear force (WBS) analysis. Mean marbling scores (n = 284) for stimulated sides did not differ (P = .923) from those for control sides within ES treatment classes. Mean values for CIE L*, a*, and b* of lean color (n = 284) were higher (P < .05) for MVMD, MVLD, HVMD, and HVLD treated sides than for the respective control sides. When WBS values for steaks were adjusted to an equal visual degree of doneness, WBS values (n = 120) were lower (P < .05) for ES treated sides than for control sides for all four types of ES application treatments. Treatment responses were not influenced by USDA Quality Grade group. For those carcasses for which the control sides had WBS values greater than 4.5 kg, matching sides treated with MVMD, MVLD, HVMD, or HVLD had WBS values less than 4.5 kg 50, 88, 60, and 75% of the time, respectively. Mean cooked steak weight loss (n = 120), adjusted to an equal visual degree of doneness, and mean purge loss (n = 24) did not differ with ES treatment.     

猪瘟病毒E2基因的克隆与鉴定

根据猪瘟病毒Brescia株全基因序列设计合成特异性引物对P1/P2对，采用异硫氰酸胍一步法（略加修改），从PK15细胞培养物中提取石门株、兔化弱毒疫苗株、野毒03株及野毒07株猪瘟病毒的总RNA。应用RT-PCR方法成功地扩增出E2全基因组约1273bp的cDNA片断，经电泳证明其大小与推测相符。分别将石门株、兔化弱毒疫苗株、野毒03株及野毒07株的E2基因片段克隆到pGEM-T载体质粒，通过对这4个重组质粒的EcoRI酶切鉴定、直接与套式PCR扩增，并对E2主要抗原区域进行224bp的序列测定，表明E2全基因克隆成功，为E2全序列测定和结构与功能的分析奠定了基础。     

Multiple forms of alkaline phosphatase in horse bone,liver, kidney,duodenum, caecum and serum separated by agarose gel electrophoresis with and without neuraminidase pretreatment

Horse bone, liver, duodenum, caecum and kidney alkaline phosphatases were separated by a commercial agarose gel electrophoresis method with and without neuraminidase pretreatment, following the manufacturer's directions. Tissue extracts were obtained in saline solution and ALP extracted from cell membranes by the butanol method. Electrophoresis was performed using a TRIS/barbital/sodium barbital buffer with detergent, pH 8.6 to 9.0, at 250 V for 30 minutes. Bone, liver and kidney untreated extracts showed two ALP bands each, but with different relative migration (compared to albumin migration). When they were preincubated with neuraminidase, the two bone bands showed a marked decrease in their migration, followed by the kidney ALP bands and the most anodic band of liver Both intestinal untreated extracts showed three bands but with different mobilities. After preincubation with neuraminidase, the three bands of caecum mucosa decreased in their migration, and the most anodic duodenum band disappeared, overlapping the second one. When tissue extracts were incubated with wheat germ-lectin (WGL), 74.5% of bone extract ALP and 67.2% of caecum extract ALP precipitated, which demonstrated that the ALP band of both tissues have similar groups in the carbohydrate side chains. Horse serum showed two electrophoretic bands, which increased to three bands when treated with neuraminidase. ALP from hepatocytes was the dominant isoform, followed by a caecum band. Because the electrophoretic mobilities of some of the tissue bands studied were identical, the neuraminidase agarose electrophoretic method appeared to be a satisfactory alternative to separate them.     

Proteolytic enzymes from Trichinella spiralis larvae.

Trichinella spiralis larvae infect their hosts by the penetration of small intestine enterocytes. The exact mechanism of penetration is unknown, but the presence of proteolytic enzymes is suspected. In this study, whole worm extracts and excretory-secretory (ES) components were obtained and their proteolytic enzymes examined. Enzymes from worm extracts were capable of hydrolysing azocoll, a general protease substrate in a wide range of pH (2-8), with maximal activity at pH 5. Trichinella spiralis larval enzymes were sensitive to metalloprotease and serine protease inhibitors. Three proteases were identified in worm extracts at molecular weight (MW) 48, 54 and 62 kDa by incorporating a gelatine substrate into a standard or a modified sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) set-up, in which we used low SDS concentration in gel and electrophoresis buffer (0.01%). Intact larvae incubated in a medium containing azocoll showed azocollytic activity. Subsequent analysis of ES products by modified SDS-PAGE in gels containing gelatine demonstrated the presence of three protease of apparent MW 33, 62 and 230 kDa.     

Osteopontin in Seminal Plasma and Sperm Membrane of Dogs

The objective of this work was to describe the presence of osteopontin (OPN) in canine seminal plasma and sperm membranes. A pool of seminal plasma and sperm membrane extract from 30 dogs was used. Polyacrylamide electrophoresis gels were performed and the bands were transferred to nitrocellulose paper and Western blot was undertaken using an antibody anti-OPN. Two and 12 bands were marked in the seminal plasma (77.2 and 15.6 kDa) and sperm membrane extracts (70.6–26.6 kDa), respectively. However, from 12 marked bands in the sperm membrane extract, only three (46.4, 37.7 and 36.5 kDa) were strongly marked. We conclude that, seminal plasma and sperm membranes from dogs contain different isoforms of OPN; yet, further studies will be necessary to determine their function in this species.     

A comparison of extracted proteins of isolates of Dermatophilus congolensis by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting.

Antigenic diversity within a collection of 18 isolates of Dermatophilus congolensis from different Continents was examined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and by Western blotting with sera from cattle with clinical dermatophilosis using whole cell extracts obtained by three methods and one extract of extracellular products of D. congolensis. One of the methods involving the release of a lysostaphin-solubilized protein (LSP) of whole cells of D. congolensis revealed a number of discrete and easily-identifiable bands in SDS-PAGE which were found suitable for characterizing protein patterns and was, therefore, subsequently used for a comparative analysis of the proteins of all the D. congolensis isolates. Six electropherotypes (ET) of D. congolensis were identified among the 18 isolates using the protein profiles based on the presence of four protein bands at Molecular weights (MW) 62, 28, 17.4 and 16.4 kDa. The ETs were found among isolates from different animal species and from different sources with ET1 consisting of three bovine and two equine isolates; ET2, two bovine and three ovine isolates; ET3, two bovine isolates; ET4, two bovine isolates; ET5, one bovine and one ovine isolates and ET6, two bovine isolates. Immunoblotting of the extracts of D. congolensis isolates with sera from cattle with clinical dermatophilosis infection demonstrated protein bands of MW ranging from 9 kDa to 188 kDa. Sera from chronic dermatophilosis infection demonstrated a 28 kDa protein which was immunodominant in the LSP extracts of all the 18 isolates of D. congolensis tested while sera from mild infections demonstrated mainly the 62 kDa protein in the same extracts. However, many protein bands were demonstrated in surface membrane (TSMP) and extracellular protein extracts with sera from only mildly infected animals. The protein patterns observed in all isolates of D. congolensis revealed global antigenic similarities and distinct differences among isolates which could not be associated with either geographic, climatic or host factors. Also sera from infected animals from endemic regions of dermatophilosis could not differentiate isolates of D. congolensis. This suggests the possibility that such sera must have come from animals that had been infected by a multitude of D. congolensis strains present in the herd environment and strains an animal could have come across during the 'ritual' annual cross-country migration of the cattle herds.