Expression conditions of human immunodeficiency virus type 2 external glycoprotein gp105 in the recombinant Pichia Pastoris strain were optimized via orthogonal test of some factors such as the rate of aeration, the i...Expression conditions of human immunodeficiency virus type 2 external glycoprotein gp105 in the recombinant Pichia Pastoris strain were optimized via orthogonal test of some factors such as the rate of aeration, the inductive duration, the initial pH and the concentration of methanol. The results from tests of between subjects effects showed that the most important parameter for efficient expression of gp105 in recombinant Pichia Pastoris strain is adequate aeration during methanol induction, and the optimum inductive condition for gp105 expression was: more than 80% aeration, 3 days for induction, th einitial pH of 6 0-7 0, the final methanol concentration of 1 0%-1 5%. With this condition, the expressed gp105 was secreted into fermentation broth and reached a ield of 30%, approximately 200 mg/L. Expressed gp105 was isolated and purified by sating out and Sephadex G 100 chromatography and the yield of gp105 was 40%. gp105 was purified to electrophoretic purity and its pI was about 5 0 by SDS PAGE and isoelectrofocusing. Its N terminal amino acid was arginine by Dansyl Cl and the result indicated that expressed gp105 was secreted and cleavaged correctly. The results from ELISA demonstrated that the purifiec gp105 showed good reactiongenicity and antigenic specificity.展开更多
本文对纯化了的猪肝DFPase~[**]进行了一系列结构和性质的测定。用重复中毒的方法证明了猪肝中低分子量二异丙基氟磷酸酯(DFP)水解酶(DFPase)是属于催化酶。分析了DFPase的氨基酸组成并求出其分子量为17900。该酶的km值为6.7×10^(-...本文对纯化了的猪肝DFPase~[**]进行了一系列结构和性质的测定。用重复中毒的方法证明了猪肝中低分子量二异丙基氟磷酸酯(DFP)水解酶(DFPase)是属于催化酶。分析了DFPase的氨基酸组成并求出其分子量为17900。该酶的km值为6.7×10^(-4)mol/L。催化水解DFP的最适pH为7.5～7.7,反应最佳温度为70℃。根据催化水解DFP、Soman的活力以及Mn^(2+)对此酶的影响证明该酶属于Mazur type DFPase~[1]。展开更多
以整合素连接激酶相关磷酸酶(ILKAP)全蛋白为抗原,采用腿部肌肉注射和耳缘静脉注射相结合的方法对新西兰大白兔进行免疫,共免疫6次,最终获得ILKAP抗血清.采用Protein A agrose柱纯化抗体,获得ILKAP的多克隆抗体.免疫印迹分析表明,所获得...以整合素连接激酶相关磷酸酶(ILKAP)全蛋白为抗原,采用腿部肌肉注射和耳缘静脉注射相结合的方法对新西兰大白兔进行免疫,共免疫6次,最终获得ILKAP抗血清.采用Protein A agrose柱纯化抗体,获得ILKAP的多克隆抗体.免疫印迹分析表明,所获得的ILKAP多克隆抗体具有较高的特异性,间接ELISA方法测定ILKAP多克隆抗体的滴度为1∶1 600.免疫荧光分析表明,所获得的ILKAP多克隆抗体在细胞水平应用效果较好.展开更多
4-Amino-dimethyl-aniline hydrochloride is one of the main components of the wastewater from vanillin production plant. So it is used for the target contamination to be degraded by microorganism, electrochemical oxidat...4-Amino-dimethyl-aniline hydrochloride is one of the main components of the wastewater from vanillin production plant. So it is used for the target contamination to be degraded by microorganism, electrochemical oxidation and bio-electro reactor in this paper. We found out a economical, efficient and non-polluted methods for the treatment of wastewater from vanillin production plant. The results indicate that there are a lot of special microbes and bacteria eating 4-amino-dimethyl-aniline hydrochloride in the activated sludge after 20 days′ incubation, at the same time, a stable bio-filmed electrode was formed. Under the condition of 25 ℃, pH=6—7. The use of bio-electro reactor not only increased the biodegradability and decreased the electrical energy consumption, but also solved the problem which the second pollution of organic compounds containing chlorine in the process of indirect electrochemical degradation on the organic pollutants.展开更多
散发性帕金森病(sporadic Parkinson's disease,sPD)的主要病理特征之一是中脑黑质致密部(substantia nigra pars compacta,SNpc)残存多巴胺能神经元内核周路易(小)体(Lewy body,LB)形成.LB发生的具体原因和确切过程有待进一步阐释...散发性帕金森病(sporadic Parkinson's disease,sPD)的主要病理特征之一是中脑黑质致密部(substantia nigra pars compacta,SNpc)残存多巴胺能神经元内核周路易(小)体(Lewy body,LB)形成.LB发生的具体原因和确切过程有待进一步阐释.来自遗传学、尸体解剖和实验科学的报道提示,蛋白酶体功能障碍及其所致的LB形成可能是按照聚集体形成途径(process of aggresomes)进行的.在聚集体形成途径过程中,异常蛋白质聚集基本上经历了非纤维化分子聚集过程(molecular crowding)以及后续的纤维化聚集过程(fibrilation of aggregation).其间,蛋白酶体功能障碍(dysfunction of proteasome)、内质网相关降解丧失(loss of endoplasmic reticulum associated degradation)、非纤维化聚集物(nonfibrilar aggregates)、聚集体(aggresomes)及至纤维化LB(fibrilar LB)等构成了sPD病变过程的主要事件.这提示在sPD病变过程中,蛋白酶体功能障碍及其所致的LB形成过程实质上是细胞信号的转导过程,其间涉及了众多的蛋白质分子.展开更多
文摘Expression conditions of human immunodeficiency virus type 2 external glycoprotein gp105 in the recombinant Pichia Pastoris strain were optimized via orthogonal test of some factors such as the rate of aeration, the inductive duration, the initial pH and the concentration of methanol. The results from tests of between subjects effects showed that the most important parameter for efficient expression of gp105 in recombinant Pichia Pastoris strain is adequate aeration during methanol induction, and the optimum inductive condition for gp105 expression was: more than 80% aeration, 3 days for induction, th einitial pH of 6 0-7 0, the final methanol concentration of 1 0%-1 5%. With this condition, the expressed gp105 was secreted into fermentation broth and reached a ield of 30%, approximately 200 mg/L. Expressed gp105 was isolated and purified by sating out and Sephadex G 100 chromatography and the yield of gp105 was 40%. gp105 was purified to electrophoretic purity and its pI was about 5 0 by SDS PAGE and isoelectrofocusing. Its N terminal amino acid was arginine by Dansyl Cl and the result indicated that expressed gp105 was secreted and cleavaged correctly. The results from ELISA demonstrated that the purifiec gp105 showed good reactiongenicity and antigenic specificity.
文摘本文对纯化了的猪肝DFPase~[**]进行了一系列结构和性质的测定。用重复中毒的方法证明了猪肝中低分子量二异丙基氟磷酸酯(DFP)水解酶(DFPase)是属于催化酶。分析了DFPase的氨基酸组成并求出其分子量为17900。该酶的km值为6.7×10^(-4)mol/L。催化水解DFP的最适pH为7.5～7.7,反应最佳温度为70℃。根据催化水解DFP、Soman的活力以及Mn^(2+)对此酶的影响证明该酶属于Mazur type DFPase~[1]。
文摘以整合素连接激酶相关磷酸酶(ILKAP)全蛋白为抗原,采用腿部肌肉注射和耳缘静脉注射相结合的方法对新西兰大白兔进行免疫,共免疫6次,最终获得ILKAP抗血清.采用Protein A agrose柱纯化抗体,获得ILKAP的多克隆抗体.免疫印迹分析表明,所获得的ILKAP多克隆抗体具有较高的特异性,间接ELISA方法测定ILKAP多克隆抗体的滴度为1∶1 600.免疫荧光分析表明,所获得的ILKAP多克隆抗体在细胞水平应用效果较好.
文摘4-Amino-dimethyl-aniline hydrochloride is one of the main components of the wastewater from vanillin production plant. So it is used for the target contamination to be degraded by microorganism, electrochemical oxidation and bio-electro reactor in this paper. We found out a economical, efficient and non-polluted methods for the treatment of wastewater from vanillin production plant. The results indicate that there are a lot of special microbes and bacteria eating 4-amino-dimethyl-aniline hydrochloride in the activated sludge after 20 days′ incubation, at the same time, a stable bio-filmed electrode was formed. Under the condition of 25 ℃, pH=6—7. The use of bio-electro reactor not only increased the biodegradability and decreased the electrical energy consumption, but also solved the problem which the second pollution of organic compounds containing chlorine in the process of indirect electrochemical degradation on the organic pollutants.
文摘散发性帕金森病(sporadic Parkinson's disease,sPD)的主要病理特征之一是中脑黑质致密部(substantia nigra pars compacta,SNpc)残存多巴胺能神经元内核周路易(小)体(Lewy body,LB)形成.LB发生的具体原因和确切过程有待进一步阐释.来自遗传学、尸体解剖和实验科学的报道提示,蛋白酶体功能障碍及其所致的LB形成可能是按照聚集体形成途径(process of aggresomes)进行的.在聚集体形成途径过程中,异常蛋白质聚集基本上经历了非纤维化分子聚集过程(molecular crowding)以及后续的纤维化聚集过程(fibrilation of aggregation).其间,蛋白酶体功能障碍(dysfunction of proteasome)、内质网相关降解丧失(loss of endoplasmic reticulum associated degradation)、非纤维化聚集物(nonfibrilar aggregates)、聚集体(aggresomes)及至纤维化LB(fibrilar LB)等构成了sPD病变过程的主要事件.这提示在sPD病变过程中,蛋白酶体功能障碍及其所致的LB形成过程实质上是细胞信号的转导过程,其间涉及了众多的蛋白质分子.
