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21.
M C Shao 《Analytical biochemistry》1992,205(1):77-82
A new rapid and sensitive method for characterizing lectin specificity using streptavidin-biotinylglycans as a tool is presented. This assay is analogous to enzyme immunoassay and takes advantage of the strong, irreversible adsorption of streptavidin to the wells of the chambers of titer plates. A series of streptavidin-biotinylglycans was first coated on a microtiter plate, and then one of six lectins, concanavalin A, wheat germ agglutinin, Phaseolus vulgaris (red kidney bean) erythro-agglutinin, Lens culinaris (lentil) agglutinin, Datura stramoniun agglutinin, or Sambucus nigra (elderberry bark) agglutinin coupled to horseradish peroxidase, was added. After incubation and thorough washing, only the lectin bound to a complementary glycan remained and could be detected and quantified by the peroxidase reaction. It was established that the lectins retained their oligosaccharide-binding specificities after coupling to the peroxidase, that the binding was inhibited by addition of the corresponding sugar inhibitors, and that the color intensity produced by the enzyme reaction is proportional to the amount of lectin-peroxidase bound to biotinylglycan complexed with streptavidin immobilized on the plate. As an example, it was found that the peroxidase-D. stramoniun agglutinin conjugate strongly bound biotinylglycans, GlcNAc3-Man5-R, GalGlcNAc3Man5-R, and GlcNAc3-4Man3-R (R = GlcNAc2-[6-(biotinamido)hexanoyl]-Asn). As little as 10 pmol/ml of lectin was detected. With the growing availability of biotinylglycans, the method should represent a reliable and simple procedure for screening lectin-oligosaccharide recognition qualitatively and quantitatively. 相似文献
22.
从嗜热蓝藻优雅粘囊藻(Myxosarcina concinna Printz.)中分离到具有放氧活性的藻胆体-类囊体膜复合物。它的吸收峰位于680、628、490、438和420 nm ,在低离子强度(0.1~0.4 m ol/L)磷酸缓冲液中的664 nm 荧光发射峰随离子强度的降低而升高,718 nm 荧光发射峰与此相反(77 。K,Ex= 580 nm )。当把游离的藻胆体和已解离去藻胆体的类囊体膜在蔗糖磷酸缓冲液中重组时,随时间延长(0~60 m in),718 nm 荧光发射峰逐渐升高,685 nm 荧光发射峰逐渐下降(77 。K,Ex = 580 nm ),表明藻胆体与类囊体之间的解离和结合对光能传递的影响 相似文献
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24.
生物复苏——大绝灭后生物演化历史的第一幕 总被引:12,自引:1,他引:11
生命史是一部生物界短期,快速剧变与长期,慢速稳定相互交替的历史。大绝灭(即集群绝灭)事件反映了全球环境的大突变,点断了地质历史中的生命记录及其发展历程,预示着生物界的演化出现了最有意义的飞跃,近年来尝试研究大绝灭后全球生物界的残存-复苏及其基本型式,并探索复苏的控制因素,标志着地质科学中一个重心的转移(即从大绝灭转向其后的生物残存与复苏的研究)。生物复苏揭示了大绝灭后生物演化历史的第一幕,其研究的 相似文献
25.
The Escherichia coli chaperonins, GroEL and GroES, as well as their complexes in the presence of a nonhydrolyzable nucleotide AMP-PNP, have been imaged with the atomic force microscope (AFM). We demonstrate that both GroEL and GroES that have been adsorbed to a mica surface can be resolved directly by the AFM in aqueous solution at room temperature. However, with glutaraldehyde fixation of already adsorbed molecules, the resolution of both GroEL and GroES was further improved, as all seven subunits were well resolved without any image processing. We also found that chemical fixation was necessary for the contact mode AFM to image GroEL/ES complexes, and in the AFM images. GroEL with GroES bound can be clearly distinguished from those without. The GroEL/ES complex was about 5 nm higher than GroEL alone, indicating a 2 nm upward movement of the apical domains of GroEL. Using a slightly larger probe force, unfixed GroEL could be dissected: the upper heptamer was removed to expose the contact surface of the two heptamers. These results clearly demonstrate the usefulness of cross-linking agents for the determination of molecular structures with the AFM. They also pave the way for using the AFM to study the structural basis for the function of GroE system and other molecular chaperones. 相似文献
26.
Micropipette suction for measuring piconewton forces of adhesion and tether formation from neutrophil membranes. 总被引:5,自引:3,他引:2
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A new method for measuring piconewton-scale forces that employs micropipette suction is presented here. Spherical cells or beads are used directly as force transducers, and forces as small as 10-20 pN can be imposed. When the transducer is stationary in the pipette, the force is simply the product of the suction pressure and the cross-sectional area of the pipette minus a small correction for the narrow gap that exists between the transducer and the pipette wall. When the transducer is moving along the pipette, the force on it is corrected by a factor that is proportional to the ratio of its velocity relative to its drag-free velocity. With this technique, the minimum force required to form a membrane tether from neutrophils is determined (45 pN), and the length of the microvilli on the surface of neutrophils is inferred. The strength of this technique is in its simplicity and its ability to measure forces between cells without requiring a separate theory or a calibration against an external standard and without requiring the use of a solid surface. 相似文献
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28.
采到骨状拟夏孢锈(Uredinopsis ossiformis Kamei)为中国新记录种。 相似文献
29.
中药凌霄花的药理学考察 总被引:2,自引:0,他引:2
实验结果表明,凌霄花对离体猪冠状动脉条具有抑制收缩的作用;对大鼠血栓形成有抑制作用;能加快红细胞电泳,增加红细胞电泳率,使血液红细胞处于分散状态;对离体孕子宫能显著增强收缩活性,增加收缩频率及增强收缩强度。以上结果可初步说明,凌霄花行血去瘀的作用及其“孕妇慎用”的合理性。凌霄花的3种混淆品(白泡桐花,毛泡桐花,兰考泡桐花)的药理作用与凌霄花不同,故不能作凌霄花使用。 相似文献
30.
利用PCR方法对单纯疱疹病毒Ⅱ型糖蛋白D(HSV-2gD)基因进行了修饰,在其5'端删去约500bp的非编码区,仅保留ATG上游7个bp。将修饰后的HSV-2gD基因插入到带有痘苗病毒天坛株TK基因区段的痘苗表达质粒pJSA1175,置于痘苗病毒P7.5k早/晚期启动子控制下。将此重组质粒用脂质体Lipofectin方法转染已受野型TK ̄+痘苗病毒天坛株感染的TK ̄-143细胞,通过同源重组机制和标志基因LacZ产物的蓝斑显色作用,以及BudR试剂对TK表型的选择压力,筛选出整合有HSV-2gD基因的重组痘苗病毒。Southem杂交表明,HSV-2gD基因已正确地插入痘苗病毒TK基因区内;间接免疫荧光检测显示,HSV-2gD蛋白已得到有效表达,且主要分布于细胞膜。重组病毒免疫家兔可产生明显的抗HSV-2gD中和抗体。用重组病毒免疫小鼠,3周后可使94%(17/18)的小鼠对抗HSV-2的致死量攻击,表明重组病毒具有明显的免疫保护作用。 相似文献