植物热激蛋白的功能及其基因表达的调控

本文介绍了植物热激蛋白的产生、分布和分类。着重论述了热激反应的特点、植物热激蛋白的功能、热激基因表达与调控的研究进展     

Hsp70与汉滩病毒S基因融合蛋白的原核表达及免疫小鼠的初步研究

本文构建了hsp70与S基因的原核融合表达载体pGEX-4T-1/hsp70-S,在大肠杆菌中表达,并通过GSTrapFF柱进行了纯化.同时制备了NP和Hsp70两种纯化蛋白.分别用这三种纯化蛋白免疫BALB/c小鼠,结果表明纯化的NP和Hsp70-NP两种蛋白均可同时诱导产生抗汉滩病毒核蛋白(NP)抗体,且后者刺激产生的抗体效价明显高于前者.淋巴细胞增殖实验表明,两组免疫小鼠的脾细胞均能够对体外抗原刺激产生增殖反应,而Hsp70-NP组免疫小鼠脾细胞对NP的增殖指数明显高于NP组免疫组.结果显示,与单独用NP免疫小鼠相比,Hsp70-NP纯化蛋白可以刺激机体产生更强的抗汉滩病毒体液免疫应答和特异性淋巴细胞增殖反应.     

Hsp70与汉滩病毒S基因融合蛋白的原核表达及免疫小鼠的初步研究

本文构建了hsp70与S基因的原核融合表达载体pGEX-4T-1/hsp70-S,在大肠杆菌中表达,并通过GSTrapFF柱进行了纯化。同时制备了NP和Hsp70两种纯化蛋白。分别用这三种纯化蛋白免疫BALB/c小鼠,结果表明纯化的NP和Hsp70-NP两种蛋白均可同时诱导产生抗汉滩病毒核蛋白(NP)抗体,且后者刺激产生的抗体效价明显高于前者。淋巴细胞增殖实验表明,两组免疫小鼠的脾细胞均能够对体外抗原刺激产生增殖反应,而Hsp70-NP组免疫小鼠脾细胞对NP的增殖指数明显高于NP组免疫组。结果显示,与单独用NP免疫小鼠相比,Hsp70-NP纯化蛋白可以刺激机体产生更强的抗汉滩病毒体液免疫应答和特异性淋巴细胞增殖反应。     

小鼠热休克蛋白gp96羧基端片段的克隆、表达及纯化

热休克蛋白(Heat shock protein)gp96(Grp94)是近年来新发现的一类糖蛋白,除了分子伴侣的功能外,现有越来越多的献报道了它在先天性免疫和获得性免疫中的重要作用。gp96可以促进抗原呈递细胞的成熟以及细胞因子的分泌。热休克蛋白抗原肽复合体可以引起特异性的细胞毒T淋巴细胞效应,应用这个特点可以设计抗病毒及抗肿瘤药物。但是gp96全长分子量大,蛋白在大肠杆菌中表达量低,不稳定,难纯化。组织提取的gp96又受组织来源和样品量的限制。对gp96的结构和功能的研究带来困难。克隆并表达了小鼠热休克蛋白gp96的羧基端560．751aa约四分之一长的功能片段,该段包含gp96的一个肽结合区和二聚化位点。将该功能片段在大肠杆菌中进行融合表达,纯化后将融合的片段切掉,并对目的片段进行了分析,结果表明该段可能是形成二聚体密切相关的片段,为进一步研究其结构和功能打下基础。     

The effects of prostaglandin A1 and prostaglandin B1 on the differentiation of cartilage in the chick embryo

Summary In organ cultures of chick embryonic limb rudiments the mean length of explants treated with 25g/ml prostaglandin B₁ (PGB₁) was significantly smaller than that of paired controls (P<0.001) after 4, 6 and 8 days in vitro. The deceleration of linear growth was constant during 8 days in vitro. Growth inhibition was confirmed by a statistically significant decrease in explant dry weight after 8 days of culture. However, PGB₁ caused no observable alteration in the histological structure of the explants. The possible role of PGB₁ in the physiological control of cartilage growth is postulated. Explants similarly treated with prostaglandin A₁ (PGA₁) at concentrations of 15 g/ml for 8 days or 20g/ml for 4 and 8 days exhibited comma and inverted commas phenomena, caused by the intermingling of chondroblasts from the epiphyseal and flattened-cell zones, which thus ceased to be distinct entities. Adenylate cyclase in the plasma membrane may be involved in this disturbance of cartilage differentiation.     

Differentiation of K562 Cells Under ELF-EMF Applied at Different Time Courses

The time-course of ELF-EMF application to biological systems is thought to be an important parameter determining the physiological outcome. This study investigated the effect of ELF-EMF on the differentiation of K562 cells at different time courses. ELF-EMF (50?Hz, 5?mT, 1?h) was applied at two different time-courses; first at the onset of hemin induction for 1?h, and second, daily 1?h for four days. While single exposure to ELF-EMF resulted in a decrease in differentiation, ELF-EMF applied everyday for 1?h caused an increase in differentiation. The effect of co-stressors, magnesium, and heat-shock was also determined and similar results were obtained. ELF-EMF increased ROS levels in K562 cells not treated with hemin, however did not change ROS levels of hemin treated cells indicating that ROS was not the cause. Overall, these results imply that the time-course of application is an important parameter determining the physiological response of cells to ELF-EMF.     

Biological stress responses to radio frequency electromagnetic radiation: are mobile phones really so (heat) shocking?

Cells phenotypically adapt to alterations in their intra- and extracellular environment via organised alterations to gene and protein expression. Many chemical and physical stimuli are known to drive such responses, including the induction of oxidative stress and heat shock. Increasing use of mobile telephones in our society, has brought focus on the potential for radio frequency (microwave) electromagnetic radiation to elicit biological stress responses, in association with potentially detrimental effects of this to human health. Here we review evidence suggesting altered gene and protein expression in response to such emissions, with particular focus on heat shock proteins. Non-thermal induction of heat shock proteins has been claimed by a number of investigations in in vitro cellular systems, and appears pleiotropic for many other regulatory events. However, many of these studies are flawed by inconsistencies in exposure models, cell types used and the independent reproducibility of the findings. Further, the paucity of evidence from in vivo experimentation is largely contradictory. Therefore, the validity of these effects in human health risk assessment remain unsubstantiated. Where possible, suggestions for further experimental clarification have been provided.     

热休克T细胞中mRNA的表达

 富集的人外周血T淋巴细胞经40—42℃保温(热休克)可诱导产生90kd和71kd的两种主要热休克蛋白(HSP),此外,62和34kd HSP也可在不同条件下诱导。从不同处理的T淋巴细胞中提取mRNA并在免网织细胞裂解液系统中进行体外转译,显示出相同的主要HSP。其中71和62kdHSP不仅在升温时还可在低温(4℃)下诱导,表明淋巴细胞中几种主要HSP的诱导机制不完全相同。比较L-~(35)S-Met参入实验和体外转译的结果提示淋巴细胞中HSP基因表达主要在转录水平调控。     

用cDNA表达阵谱分析小鼠鼻咽部相对特异表达基因

利用Mouse Atlas^TM cDNA expression array检测鼻咽、气管、食管、膀胱四种组织中588个已知基因的表达谱,得到11个在鼻咽上皮中表达相对较高的基因,作为鼻咽部组织相对特异基因的候选者,并用RT-PCR进一步验证.     

Expression of gamma-tubulin during the development of nematode <Emphasis Type="Italic">Caenorhabditis elegans</Emphasis>

Gamma-tubulin is a centrosomal protein found in microtubule organizing centres (MTOCs) in cells from many different organisms, and has several properties, which makes it a candidate for both the initiation of microtubule assembly and anchorage. Gamma-tubulin is encoded by a single gene tbg-1 in Caenorhabditis elegans. In this paper tbg-1 was studied to understand the essential role of gamma-tubulin in C. elegans. Essential role of tbg-1 expression was confirmed by the disruption of the gene expression by gamma-tubulin anti-sense RNA production in vivo under the heat shock promoter that caused lethality in the nematodes. Expression of tbg-1 deduced from Northern blot analysis during the development revealed differential expression in different developmental stages. Using tbg-1::lacZ fusion gene expression studies in the germ line transformed worms, it was further revealed that gamma-tubulin expression was observed through out the development from embryonic and post-embryonic stages.