Synthesis of artificial antigens of diethylstilbestrol and preparation of its antibody

Diethylstilbestrol-hemi-succinate (DES-HS) and diethylstilbestrol-monocarboxylic-propyl-ether (DESMCPE) were synthesized in different ways. The two kinds of haptens were conjugated to bovine serum albumin (BSA) and ovalbumin (OVA) to form the immuno-antigens and the coating-antigen, respectively. The antibody with high titer was obtained after immunising rabbits. The results showed that the antibody had a high affinity to diethylstilbestrol tested by enzyme-linked immunosorbent assay (ELISA) and Western blot. This study lays a basis for further research on the immune test kit for the DES remnant.     

鸡卵清蛋白基因调控序列指导人促红细胞生成素转基因鸡的制备

本研究将鸡卵清蛋白5端调控区调控人促红细胞生成素的pOV-hEPO特异表达载体用脂质体包埋，和供体细胞融合后，显微注射入450枚受体胚盘，采用代用蛋壳法培养制备转基因鸡，PCR和斑点杂交检测外源基因已整合入早期鸡胚胎生殖系统，为制备表达人促红细胞生成素的鸡输卵管生物反应器奠定了基础。     

双抗夹心酶联免疫分析法检测食品中鸡蛋过敏原

[目的]建立食品中鸡蛋过敏原的酶联免疫检测方法。[方法]以卵白蛋白为检测对象,分别制备抗卵白蛋白的多克隆和单克隆抗体。将单克隆抗体作为包被抗体,辣根过氧化物酶标记多克隆抗体为二抗构建卵白蛋白的双抗夹心ELISA方法。[结果]该方法的IC50为260 ng/ml,检出限为10 ng/ml,样品的添加回收率为84.2%～89.8%,相对标准偏差小于10%。[结论]该试验建立的分析方法稳定可靠,能满足食品中鸡蛋过敏原准确、快速的检测需要。     

Induction of ovalbumin synthesis in immature chicks by actinomycin D and thioacetamide

Actinomycin D and thioacetamide induced ovalbumin synthesis and increased serum progesterone concentrations in immature chicks. The increase in progesterone induced by the carcinogens actinomycin D and thioacetamide may account for the induction of ovalbumin synthesis.     

Protein synthesis: differential stimulation of cell-specific proteins in epithelial cells of chick oviduct

Immunofluorescent and radioautographic studies demonstrate that ovalbumin and avidin are cell-specific proteins synthesized by different epithelial cells in the chick oviduct mucosa. The mechanism of the selective induction of ovalbumin synthesis by estrogen and of avidin synthesis by progesterone may be through stimulation of specific target cells by these hormones.     

鸡卵清蛋白基因调控序列的克隆与载体构建

卵清蛋白（ovalbumin）基因在鸡基因组中只有一对等位基因,却能每天合成分泌多达2 g的蛋白,占据卵白蛋白质的50%以上,是外源基因载体表达调控构件的首选。该研究从输卵管特异性启动子方面着手,通过对卵清蛋白基因启动子的筛选优化,找出启动子增强因子的位置以及组织特异性区域。将卵清蛋白基因上游-922～-2 073和-2 801～-3 100区域平均分成12个长度约为150 bp的序列,分别插入到-921～+38序列的上游,成功构建12个系列表达载体,为进一步筛选短缩版优化启动子提供材料;卵清蛋白基因第一内含子区域截断成300 bp左右的迷你内含子序列,成功构建8个迷你内含子系列载体,为筛选优化的迷你内含子提供必要的材料;成功分离鸡输卵管上皮细胞并优化电转染条件,通过荧光素酶活性检测初步筛选出具有最强活性重组质粒pGL4 UP 1412和内含子重组质粒pGL4 mini intron 3,同时推断出若干包含增强子序列区域。     

糖基化对卵白蛋白分子特性及乳化性的影响

 【目的】对糖基化卵白蛋白分子特性及乳化性进行研究,探讨二者之间内在联系。【方法】研究糖基化进程中游离氨基含量、Zeta电位、疏水性、乳化液滴平均粒径（d32）与乳化活性的变化情况。在此基础上对以上因子进行相关性分析,并对糖基化卵白蛋白的乳化液进行显微镜观察。【结果】糖基化反应使游离氨基降低41.56%、Zeta电位降低20 mV、乳化液滴平均粒径（d32）减少2.0μm,而疏水性和乳化活性分别提高2.3倍和3.9倍。各因子间明显的相关性表明,所研究因子为影响乳化性的关键因子。显微镜观察结果表明,糖基化卵白蛋白易于形成较小的乳化液滴。【结论】糖基化是通过改变蛋白质表面电荷和形成双亲结构来改善乳化性的。     

鸡蛋清中3种蛋白质的连续化提取工艺初步研究

研究鸡蛋清中溶菌酶、卵转铁蛋白和卵白蛋白的分离提取工艺.采用硫酸铵沉淀和离子交换层析分离3种蛋白质,并用聚丙烯酰胺凝胶电泳(SDS-PAGE)检测其纯度.结果显示,层析纯化后SDS-PAGE检测其纯度达到电泳纯,溶菌酶的比活由144.13 U/mg提高到2 235 U/mg,纯化倍数达15倍,回收率为15.76％;卵转铁蛋白抑菌率为48.84％.该方法分离溶菌酶、卵转铁蛋白和卵白蛋白的工艺路线简单、快捷、成本低,适用于工业生产.     

甲拌磷人工抗原的合成及多克隆抗体制备

以五硫化二磷与无水乙醇为原料,首先制得硫化物中间体,再添加甲醛和2-巯基乙醇进行反应,得到甲拌磷半抗原.在DMAP作为催化剂的条件下,甲拌磷半抗原与丁二酸酐反应,得到甲拌磷半抗原的衍生物.通过碳二亚胺法将甲拌磷半抗原衍生物分别与牛血清白蛋白(BSA)和鸡卵清白蛋白(OVA)共价偶联,得到免疫抗原和包被抗原.用免疫抗原免疫兔子获得了高效价的多克隆抗体,抗血清效价达到了1∶50 000.通过试验确立甲拌磷标准曲线,检测限为6.383μg/L,检测线性范围为6.383～10 000μg/L.     

Cytochalasin B inhibits lymphotoxin production by antigen-stimulated lymphocytes

Lymph node cells of rats sensitized with hen ovalbumin produced lymphotoxin after 6 to 12 hours of exposure to specific antigen. Lymphotoxin was assayed by its cytotoxicity for fibroblasts from syngeneic embryos during a 72-hour incubation. Cytochalasin B inhibited lymphotoxin production, as well as later DNA synthesis, at concentrations (0.1 to 5.0 micrograms per milliliter) comparable to those which affect microfilament function and cell motility in other systems, and this inhibition was reversible. Binding of antigen was not affected.     

鸡蛋清中3种蛋白质的连续化提取工艺初步研究

[目的]研究鸡蛋清中溶菌酶、卵转铁蛋白和卵白蛋白的分离提取工艺。[方法]采用硫酸铵沉淀和离子交换层析分离三种蛋白质,并用聚丙烯酰胺凝胶电泳（SDS-PAGE）检测其纯度。[结果]层析纯化后 SDS-PAGE检测其纯度达到电泳纯,溶菌酶的比活由144.13 U/mg提高到2235 U/mg,纯化倍数达15倍,回收率为15.76%;卵转铁蛋白抑菌率为48.84%。[结论]该方法分离溶菌酶、卵转铁蛋白和卵白蛋白,工艺路线简单、快捷、成本低,适用于工业生产。     

湿法糖基化改善卵白蛋白的起泡性

采用湿法糖基化改性以改善卵白蛋白的起泡性,考察葡萄糖添加量、pH、温度和时间对卵白蛋白起泡性及其稳定性的影响,在单因素试验的基础上通过响应面法优化改性工艺条件.结果表明,经湿法糖基化改性后卵白蛋白的起泡性得以显著提高,最佳改性工艺条件为:反应温度60℃、pH 7.4、葡萄糖添加量3.5%、加热时间30 min.此条件下改性后卵白蛋白的起泡性为168%,与未改性卵白蛋白起泡性(120%)相比,其起泡性提高40%.     

基于皮克林乳液的低芥酸菜籽油基凝胶油的性质及应用

以低芥酸菜籽油为基料油,结合食品级的卵清蛋白、黄原胶与单宁酸,通过高速剪切制备成具有塑性的皮克林乳液凝胶油,研究添加黄原胶与单宁酸后对皮克林乳液凝胶油性质的影响,并探究该皮克林乳液凝胶油制作的蛋糕的性质.结果表明:最高可制备出油相达70％的皮克林乳液凝胶油,形成的乳液表现为具有黏弹性的凝胶状半固体;黄原胶和单宁酸的加入...     

鸭卵清蛋白5'端调控区的克隆和序列分析

通过PCR技术从产蛋鸭输卵管基因组中扩增出1．2kb的鸭清蛋白5’端调控区,将其亚克隆入phDl8-T载体的多克隆位点(命名为pOV),经酶切和测序鉴定可作为启动外源基因表达的调控序列。为构建其启动外源基因的质粒表达载体作准备。     

磺胺间甲氧嘧啶全抗原的制备研究

采用重氮化法，将半抗原磺胺间甲氧嘧啶（SMM）与载体牛血清白蛋白（BSA）和卵清白蛋白（OVA）偶联，制得磺胺间甲氧嘧啶全抗原SMM—BSA和SMM—OVA。通过其紫外扫描光谱、SDS—PAGE凝胶电泳试验，表明半抗原和载体成功偶联，免疫动物以后可以产生满意效价。这另进一步制备抗SMM抗体提供了良好的免疫原。     

Evaporation enhancement by protein films

Evaporation of water takes place more rapidly from buffered solutions of ovalbumin or hemoglobin than from solutions of certain surface-active agents or from buffer alone. The effect appears to be connected with adsorption of protein.     

桐子叶水提物对小鼠的抗敏作用研究

[目的]研究桐子叶水提物对小鼠的抗敏作用。[方法]用卵清蛋白作为致敏抗原,制作小鼠致敏模型,豌察桐子叶水提物对小鼠过敏反应的作用。[结果]高、中浓度组桐子叶水提物对卵清蛋白引起的小鼠过敏反应均有明显的抑制作用。[结论]桐子叶水提物对卵清蛋白致敏的小鼠有明显的抗敏作用。     

Macrophage spreading: inhibition in delayed hypersensitivity

The capacity of peritoneal macrophages to spread was studied with cells of mice infected with Listeria monocytogenes and with cells of guinea pigs sensitized with BCG (bacille Calmette Guérin) vaccine or immunized with ovalbumin. In macrophages taken from animals having delayed hypersensitivity, this ability was markedly decreased by the presence of specific antigen for less than 1 hour. Such an effect was not observed in guinea pigs having only circulating antibodies.     

Specific immunologic unresponsiveness to synthetic polypeptide antigens

The injection of two synthetic polypeptide antigens into adult rabbits treated with 6-mercaptopurine, or into newborn rabbits, resulted in immunological "unresponsiveness" to subsequent immunizations with these antigens. The "tolerant" animals were shown to be reactive toward the non-related antigen ovalbumin.