förster resonance energy transfer measurements and their use in structura

... C-terminal citrine) and citrine–GAPDH (chimeric protein with N-terminal citrine and C-terminal GAPDH) were localized in the cytoplasm PGK–cerulean (chimeric protein with N-terminal PGK and C-terminal cerulean) ... finding indicates that induction of FRET does not simply depend on the length of linkers In the linker containing seven amino acids, a Pro-Pro sequence that is not contained in other linkers is incorporated ... GAPDH and PGK linked to cerulean or citrine were introduced into CHO-K1 cells As shown in Figs 1A and S1A, transiently expressed GAPDH–citrine (chimeric protein with N-terminal GAPDH and C-terminal...

Ngày tải lên: 16/02/2014, 09:20

... target, and the selection of aptamers with both fast and slow binding kinetics It also can be used to accurately determine the binding affinities, kinetics and specificity of aptamer sequences In ... for 12 partitioning include biotin-streptavidin, glutathione S-transferase (GST), polyhistidine tagging and Immobilized metals49; 50 Biotin-streptavidin was originally used for affinity column based ... the use aptamers as affinity probes in ACE include IgE, HIV type reverse transcriptase, thrombin and antithrombin III12-14 The use of aptamers in biosensors is becoming a huge area of interest in...

Ngày tải lên: 10/09/2015, 09:22

... as the metal binding event would be totally independent of the interactions involved in the enzyme–substrate complex The binding of MUG and Mg2+ are not independent, however, but instead are highly ... donor involved in the transferase Fig Diagram showing the thermodynamic equilibria and their dissociation constants involved in the binding of UDP-Glc, MUG and Mg2+ to TcdA Values shown in parentheses ... spectroscopic handles that can be used to probe directly the Mg2+ binding to TcdA [29] Mg2+ binding to free TcdA is invisible to our assays Thus, in this study, we have resorted to looking at the...

Ngày tải lên: 08/03/2014, 22:20

... herefrom, that: 11 In polar coordinates, the sine-wave is represented in intensity and phase by a vector o c, and in combining or dissolving sine-wa/aes, they are to be combined or dissolved by ... space, and of which the complex quantity in a plane is only a special case M B STKINIHTTZ: In introducing j , first as distinguishing index and then defining it as V — my object was to introduce ... 01 — 05,/ Case Non-inductive line of Non-inductive load of Case Non-inductive line of Inductive load of Case Inductive line of Inductive load of Case Inductive line of Non-inductive load of 1...

Ngày tải lên: 04/06/2014, 12:33

... indicators and their use in sustainable agriculture and rural development 19 Land quality indicators and their use in sustainable agriculture and rural development 19 The context of indicators in FAO ... issues and participatory approaches Land quality indicators and their use in sustainable agriculture and rural development 11 CONCEPTS, DEFINITIONS AND PRINCIPLES "Land", the "functions of land", ... examples of improvements in land quality (e.g., results of soil conservation and land -use planning case studies); Land quality indicators and their use in sustainable agriculture and rural development...

Ngày tải lên: 06/07/2014, 08:20

... differences in the techniques used in different laboratories are minor ones, principally concerning the use or not of an automatic sequencing machine for size determination For SNP genotyping, this ... families for genotyping, selecting recombination events in recurrent backcrosses, using advanced intercross lines (AIL) or performing linkage disequilibrium and haplotype-based studies in outbread populations ... just using SNPs in animal genetics 277 standard oligonucleotides in combination (in addition to restriction enzymes in the case of AFLPs), considerably reducing the effort and consumables, and therefore...

Ngày tải lên: 09/08/2014, 18:21

Ngày tải lên: 14/11/2014, 08:12

... described below, thereby showing it has attained its native structure This purified protein showed single and double stranded DNA binding activity Binding to single stranded DNA stimulated a significant ... annealing of FRET dye labeled strands (Phi-W and Phi-C; Fig 1B) Interestingly enough, the three stranded pairing reaction exhibited OsDmc1 concentration dependent increase in fluorescein emission intensity ... presynaptic complex (containing Phi-W strand) was incubated in the absence of Phi-C strand, showed no loss in fluorescence intensity as a function of time (Fig 2A, line 1), thereby revealing that the rapid...

Ngày tải lên: 19/02/2014, 07:20

... Jockers, R (2002) Monitoring of ligand-independent dimerization and ligand-induced conformational changes of melatonin receptors in living cells by bioluminescence resonance energy transfer J Biol Chem ... limitations, for example changes in energy transfer observed could be caused by conformational changes in the proteins rather than changes in protein– protein interaction A combination of several methods ... resuspended in buffer and stored at )80 °C in aliquots of 500 lL The protein concentration was determined by the method of Lowry et al [32], using BSA as the standard Radioligand-binding assay...

Ngày tải lên: 21/02/2014, 00:20

... results obtained using cis-PnA as a fluorescent reporter, indicate that the increase in the cis-PnA fluorescence intensity was due to its incorporation into the NaTDC micelles, resulting in a drastic ... lids in both molecules (A and B) are well defined in the electron density maps and are in the same fully opened conformations They are not involved in the interactions between molecule A and B, and ... TLL The stereoview of OA binding modes in molecule A and B (OA_A and OA_B, respectively, thick lines); C9 and C10 correspond to the position of these atoms in the OA_A ligand, where the electron...

Ngày tải lên: 08/03/2014, 23:20

... encoding: (C) cerulean-elongin B and elongin Ccitrine; (D) elongin B-cerulean and elongin C-citrine; (E) ceruleanelongin B and citrine-elongin C; and (F) elongin B-cerulean and citrine-elongin ... cerulean-linked elongin B and citrine-linked elongin C can be detected in the following donor-acceptor combinations in decreasing order: cerulean-elongin B and citrine-elongin C > cerulean-elongin B and ... elongin C Results Imaging of interaction between elongin B and elongin C As shown in Fig 1A, cerulean-elongin B and elongin B-cerulean were expressed throughout cells, and citrine-elongin C and...

Ngày tải lên: 16/03/2014, 05:20

... as in (A) cAMP accumulation in the transfected HEK 293 cells was measured for after adding, 20 lM forskolin and 10 lM prostaglandin E1 for transfections involving AC5 and AC6; 20 lM forskolin, ... camera (Princeton Instruments), z-step motor and dual ®lter wheels controlled by SLIDEBOOK 3.0 software (Intelligent Imaging Innovation, Denver, CO, USA) Binning ´ mode and 500 ms integration ... calmodulin, binding to the C-terminus [22] Issues of speci®city of intermolecular interactions were addressed with AC5 or AC6, which are inhibitable by Ca2+, apparently independently of calmodulin...

Ngày tải lên: 23/03/2014, 21:21

... containing CCh, which induced a second calcium increase and a further increase in Ins(1,4,5)P3 (Fig 3Aa) Both of these calcium increases were accompanied by increased Ins(1,4,5)P3, indicating ... fluorescence intensity Increases in cellular calcium and Ins(1,4,5)P3 were induced using carbachol (CCh; 100 lm), which specifically activates mAChR and induces Ins(1,4,5)P3 production in PC12 cells, ... Calcium-dependent enhancement of receptorinduced Ins(1,4,5)P3 increases in CHO cells expressing m1 mAChR PC12h cells express m1 mAChR and CCh-induced increases in calcium and investigate the cell-type dependence...

Ngày tải lên: 30/03/2014, 03:20

... HEK 293 cells were used instead of tsA cells Fluorescence and luminescence measurements For the measurements of fluorescence and luminescence in cells cotransfected with Rluc- and GFP2-constructs, ... oligomerization using time-resolved fluorescence resonance energy transfer and bioluminescence resonance energy transfer J Biol Chem 276, 14092–14099 Xu, Y., Piston, D.W & Johnson, C.H (1999) A bioluminescence ... following day, the cells were washed three times in NaCl/Pi, resuspended in 300 lL NaCl/Pi and distributed in black optiplates (Packard) Fluorescence and luminescence recordings were performed in...

Ngày tải lên: 08/03/2014, 09:20

Ngày tải lên: 24/03/2014, 00:20

... and Dr Randolph Stone for helping me to gain some degree of insight into the workings of the four elements, and also to the many friends and students who have encouraged my writing and teaching ... Knowledge L7 "Beginner'smind" is our original mind, actually an empty and ready mind If our mind is empty, it is always ready for anymind there are thing; it is opento everything .In the beginner's many ... of "thinking" and "intuition" to indicate the two groups.) The thinking faculty functions through the systematic classification and discrimination of facts which are then arranged in certain patterns...

Ngày tải lên: 29/03/2014, 04:20

... method you use to get what you want: Mars in air uses persuasion, Mars in fire uses power and initiative, Mars in earth uses patience and efficiency, and Mars in water use slyness,intuition, and a ... deeperunderstanding of the psychological dynamics involved, but it is also useful for gaining insight into how people effect each other's health simply by their being together Even if we are involved in ... man with the Sun in Virgo and Moon in Leo, having no major or minor aspect between them The Sun and Moon in this example are in incompatible elements; and, al- P otential for Integration 131 though...

Ngày tải lên: 29/03/2014, 04:20

... events in the main proceedings and that those measures designate as navigable waters the waters in which the accused in the main proceedings used personal watercraft and consequently had proceedings ... waters and waterways have the effect of preventing users of personal watercraft from using them for the specific and inherent purposes for which they were intended or of greatly restricting their use, ... affixing tinted film, potential customers, traders, or individuals had practically no interest in buying tinted film.174 In Commission v Italy, the Grand Chamber spoke of the prohibition having...

Ngày tải lên: 30/03/2014, 06:20

... efficient and convenient method for measuring protein– protein interaction The same insect cell line can be routinely used to express any recombinant proteins of interest, allowing various combinations ... detect protein–protein interactions and protein conformational changes in vivo Curr Opin Struct Biol 11, 573– 578 20 van Thor, J.J & Hellingwerf, K.J (2002) Fluorescence resonance energy transfer ... optimization The assay was optimized in order to determine the optimal infection intensity for individual recombinant protein expression and conditions of measuring FRET Protein expression was optimized...

Ngày tải lên: 30/03/2014, 20:20

... the older ones, scorched and dying at tip and outer margins Leaf margins often crinkled and curled Corn stalks may be brittle (browning of tissue evident in split joints, especially toward base ... present, generally within interveinal tissue surrounding midrib, veins remain green, younger leaves yellow ish (striping in grasses) or even white (white bud in corn) ZINC 11 Fertilizer Materials ... Calculating Nutrient Content of Dry and Liquid Fertilizers Frequency of Fertilizer and Lime Applications Timing of Fertilizer and Lime Applications Application Methods Broadcasting Banding Sidedressing...

Ngày tải lên: 20/04/2014, 16:10