CLC Chloride Channels in Caenorhabditis elegans*
... encodes six putative chloride channels (CeCLC-1 through CeCLC-6) that represent all three known branches of the mammalian CLC gene family. Using promoter fragments to drive the expression of the green fluorescent protein, CeCLC-2, -3, and -4 expression was studied in transgenic C. elegans. CeCLC-4 w ...
... encodes six putative chloride channels (CeCLC-1 through CeCLC-6) that represent all three known branches of the mammalian CLC gene family. Using promoter fragments to drive the expression of the green fluorescent protein, CeCLC-2, -3, and -4 expression was studied in transgenic C. elegans. CeCLC-4 w ...
Kinesin Microtubule Gliding Assay 1. Make labeled microtubules
... 1) If you have a very slow moving motor it can be useful to flow in just MTs diluted in BCB and allow them to bind (turn coverslip face down and wait a few minutes). Then flow in motility mix containing everything except MTs. This will reduce the background fluorescence and prevent extra MTs binding ...
... 1) If you have a very slow moving motor it can be useful to flow in just MTs diluted in BCB and allow them to bind (turn coverslip face down and wait a few minutes). Then flow in motility mix containing everything except MTs. This will reduce the background fluorescence and prevent extra MTs binding ...
7 Putting genetics into optogenetics: knocking out proteins with light
... The RING finger E3 ligase family has been used to destabilize specific proteins [8]. These proteasome-dependent degrons differ in their method for inducing interactions between the E3 ubiquitin ligase complex and the target protein. The protein can be destabilized by: amino acids at the N-terminus o ...
... The RING finger E3 ligase family has been used to destabilize specific proteins [8]. These proteasome-dependent degrons differ in their method for inducing interactions between the E3 ubiquitin ligase complex and the target protein. The protein can be destabilized by: amino acids at the N-terminus o ...
Dissecting the function of Atg1 complex in Dictyostelium autophagy
... AreA to determine if these proteins localize to autophagosomes. For this, the different proteins were tagged with GFP, expressed in wild-type Ax4 cells and analysed by confocal microscopy (figure 2). We found that Atg101, FIP200, Atg13 and AreA are uniformly distributed in the cytoplasm. None of the ...
... AreA to determine if these proteins localize to autophagosomes. For this, the different proteins were tagged with GFP, expressed in wild-type Ax4 cells and analysed by confocal microscopy (figure 2). We found that Atg101, FIP200, Atg13 and AreA are uniformly distributed in the cytoplasm. None of the ...
Blockade of Stimulus Convergence in Amygdala Neurons Disrupts
... manipulation of CREB (cAMP response element-binding protein) levels, and reversible inactivation of targeted cells to selectively block stimulus convergence (Zhou et al., 2009). Recent studies have suggested that amygdala neurons with relatively high CREB levels are preferentially recruited in assoc ...
... manipulation of CREB (cAMP response element-binding protein) levels, and reversible inactivation of targeted cells to selectively block stimulus convergence (Zhou et al., 2009). Recent studies have suggested that amygdala neurons with relatively high CREB levels are preferentially recruited in assoc ...
Distinct Lytic Vacuolar Compartments are Embedded
... native promoter Based on the observation of TIP1;1::GFP fusion protein fluorescence, Hunter and co-workers reported that TIP1;1 is not present in Arabidopsis seedlings during early seed germination and appears only 3.5 d after germination when it replaces a-TIP (Hunter et al. 2007). TIP1;1 is the ma ...
... native promoter Based on the observation of TIP1;1::GFP fusion protein fluorescence, Hunter and co-workers reported that TIP1;1 is not present in Arabidopsis seedlings during early seed germination and appears only 3.5 d after germination when it replaces a-TIP (Hunter et al. 2007). TIP1;1 is the ma ...
Glycan-independent Role of Calnexin in the Intracellular
... buffer. Immunocomplexes were released by boiling for 5 min in SDS sample buffer and separated by SDS-PAGE on a 10% gel. Immunofluorescence and Photobleaching Experiments—For immunofluorescence microscopy, cells were fixed with 3% paraformaldehyde in phosphate-buffered saline for 20 min at room tempe ...
... buffer. Immunocomplexes were released by boiling for 5 min in SDS sample buffer and separated by SDS-PAGE on a 10% gel. Immunofluorescence and Photobleaching Experiments—For immunofluorescence microscopy, cells were fixed with 3% paraformaldehyde in phosphate-buffered saline for 20 min at room tempe ...
Bioluminescence Microscopy
... Advantages and Challenges In bioluminescence microscopy we detect light that is produced due to a chemical reaction of an enzyme (luciferase) with its substrate (luciferin). Similar to the much better known fluorescence approaches, bioluminescence is a technique that can be used for non-invasive ana ...
... Advantages and Challenges In bioluminescence microscopy we detect light that is produced due to a chemical reaction of an enzyme (luciferase) with its substrate (luciferin). Similar to the much better known fluorescence approaches, bioluminescence is a technique that can be used for non-invasive ana ...
File
... With reference to Table 4.2, and the information in (iii), state the cause or causes of reduced fertility in these mutant worms at each temperature. ...
... With reference to Table 4.2, and the information in (iii), state the cause or causes of reduced fertility in these mutant worms at each temperature. ...
Protection of Drosophila chromosome ends through minimal
... Fig. 1. K81 and HipHop show a complementary localization at telomeres in the male germline. (A) Schematic representation of Drosophila spermatogenesis. Spermatogenesis is organized in differentiating units called cysts that progress along the axis of the testis (Fuller, 1993). At the apical end of t ...
... Fig. 1. K81 and HipHop show a complementary localization at telomeres in the male germline. (A) Schematic representation of Drosophila spermatogenesis. Spermatogenesis is organized in differentiating units called cysts that progress along the axis of the testis (Fuller, 1993). At the apical end of t ...
Ubiquitin-Proteasome Dependent Regulation of
... 2012). Hence, we tested the effect of Suc on the accumulation of GLK1. We extracted crude proteins from wild-type plants grown on MS plates containing different concentrations of Suc. To control for osmotic pressure, we also included plants grown on mannitol. Wild-type plants accumulated only small ...
... 2012). Hence, we tested the effect of Suc on the accumulation of GLK1. We extracted crude proteins from wild-type plants grown on MS plates containing different concentrations of Suc. To control for osmotic pressure, we also included plants grown on mannitol. Wild-type plants accumulated only small ...
The transcriptional regulator CprK detects chlorination by combining
... [17,22]. To avoid any effects coupled to the redox state of the cell, we used a CprK C200S mutant as the basis for all our studies (referred to hereafter as CprK), as it was previously shown that oxidative disulfide bond formation involving C200 inactivates the regulator [23]. A region comprising 12 ...
... [17,22]. To avoid any effects coupled to the redox state of the cell, we used a CprK C200S mutant as the basis for all our studies (referred to hereafter as CprK), as it was previously shown that oxidative disulfide bond formation involving C200 inactivates the regulator [23]. A region comprising 12 ...
Monomeric alpha-catenin links cadherin to the actin cytoskeleton
... Ridhdhi Desai1,3 , Ritu Sarpal1,3 , Noboru Ishiyama2 , Milena Pellikka1 , Mitsuhiko Ikura2 and Ulrich Tepass1,4 The linkage of adherens junctions to the actin cytoskeleton is essential for cell adhesion. The contribution of the cadherin–catenin complex to the interaction between actin and the adhere ...
... Ridhdhi Desai1,3 , Ritu Sarpal1,3 , Noboru Ishiyama2 , Milena Pellikka1 , Mitsuhiko Ikura2 and Ulrich Tepass1,4 The linkage of adherens junctions to the actin cytoskeleton is essential for cell adhesion. The contribution of the cadherin–catenin complex to the interaction between actin and the adhere ...
Identification of proteins that putatively bind the
... based on turgor pressure-induced changes in their shape. Stomatal aperture and density are affected by environmental stimuli such as light quality and quantity, CO2 concentrations, and water availability. The basic helix-loop-helix (bHLH) transcription factor MUTE drives the differentiation of trans ...
... based on turgor pressure-induced changes in their shape. Stomatal aperture and density are affected by environmental stimuli such as light quality and quantity, CO2 concentrations, and water availability. The basic helix-loop-helix (bHLH) transcription factor MUTE drives the differentiation of trans ...
letters - Cytomorpholab
... indicates that AtFH5 has a role in cytokinesis. A fluorescent dotted pattern of undetermined identity was also observed in the cytoplasm (Fig. 3) and might not reflect the true localization of AtFH5, as potential artifacts may be associated with overexpression of AtFH5–GFP. The endosperm in the Arab ...
... indicates that AtFH5 has a role in cytokinesis. A fluorescent dotted pattern of undetermined identity was also observed in the cytoplasm (Fig. 3) and might not reflect the true localization of AtFH5, as potential artifacts may be associated with overexpression of AtFH5–GFP. The endosperm in the Arab ...
Electrostatic Interactions in Wild
... reconstituted with the chlorophyll derivative zinc pyrochlorophyllide a. Absorption and fluorescence spectra were quite similar for wild-type and all mutant proteins reconstituted with this derivative. These results do not support the point charge model for the red shifts observed in the spectra of ...
... reconstituted with the chlorophyll derivative zinc pyrochlorophyllide a. Absorption and fluorescence spectra were quite similar for wild-type and all mutant proteins reconstituted with this derivative. These results do not support the point charge model for the red shifts observed in the spectra of ...
binding domains demonstrated in a plant split
... exist in a concentration-dependent equilibrium between the monomeric and dimeric state (Reddick et al., 2007; Yeh et al., 2007). This observation and the crystal structures available for Arabidopsis and pea Toc33 indicate the formation of stable homodimers of the G-domain (Sun et al., 2002; Koenig e ...
... exist in a concentration-dependent equilibrium between the monomeric and dimeric state (Reddick et al., 2007; Yeh et al., 2007). This observation and the crystal structures available for Arabidopsis and pea Toc33 indicate the formation of stable homodimers of the G-domain (Sun et al., 2002; Koenig e ...
A novel meiosisspecific protein of fission yeast, Meu13p, promotes
... loci and centromeres are sketched in insets. Crosses indicated as D/D and +/+ correspond to the cross between Dmeu13 strains (NP32-2C and -2D) and between wild-type strains (NP32-2A and -2B), respectively. Of these two cross types, 210 and 292 tetrads were analysed, respectively, of which 157 and 16 ...
... loci and centromeres are sketched in insets. Crosses indicated as D/D and +/+ correspond to the cross between Dmeu13 strains (NP32-2C and -2D) and between wild-type strains (NP32-2A and -2B), respectively. Of these two cross types, 210 and 292 tetrads were analysed, respectively, of which 157 and 16 ...
Lysosomal biogenesis and function is critical for necrotic cell death
... enhancing the activity of the low pH–dependent proteases that dismantle the cell (Syntichaki et al., 2005). To emulate impaired lysosomal acidification in degenerating neurons, we treated animals expressing a neurotoxic gain-of-function (gf ) mec-4(d) allele encoding a hyperactive ion channel subuni ...
... enhancing the activity of the low pH–dependent proteases that dismantle the cell (Syntichaki et al., 2005). To emulate impaired lysosomal acidification in degenerating neurons, we treated animals expressing a neurotoxic gain-of-function (gf ) mec-4(d) allele encoding a hyperactive ion channel subuni ...
The term fluorescence was coined by Stokes circa 1850 to name a
... It is worth noting that almost all currently used calcium-sensitive dyes were designed in the laboratory of Dr. Roger Tsien who received Nobel Prize in 2008 for his crucial contribution in the understanding and development of the green fluorescent protein, GFP. Shall we say that his prior contributi ...
... It is worth noting that almost all currently used calcium-sensitive dyes were designed in the laboratory of Dr. Roger Tsien who received Nobel Prize in 2008 for his crucial contribution in the understanding and development of the green fluorescent protein, GFP. Shall we say that his prior contributi ...
Capturing protein interactions in the secretory pathway of living cells
... although it was correctly expressed at similar levels as MCFD2YFP2 (Fig. 3 B and D). The results demonstrate the selectivity and specificity of YFP PCA to detect luminal protein–protein interactions in the secretory pathway. Moreover, coexpression of YFP1-p53 with MCFD2-YFP2 showed similar fluoresce ...
... although it was correctly expressed at similar levels as MCFD2YFP2 (Fig. 3 B and D). The results demonstrate the selectivity and specificity of YFP PCA to detect luminal protein–protein interactions in the secretory pathway. Moreover, coexpression of YFP1-p53 with MCFD2-YFP2 showed similar fluoresce ...
8. lecture
... Focusing the beam decreases its size but increases its intensity and results in not constant light absorption across the cuvette: the absorption will be the largest in the focal point where the incident intensity is the highest. As the excitation is strongly localized in a small spot at the focal po ...
... Focusing the beam decreases its size but increases its intensity and results in not constant light absorption across the cuvette: the absorption will be the largest in the focal point where the incident intensity is the highest. As the excitation is strongly localized in a small spot at the focal po ...
Protein splicing of PRP8 mini-inteins from species of the genus
... RGSHHHHHH-tag in case of the prp8-ir. The PCR amplification products containing the prp8 intein coding sequence with flanking extein sequences (5aa and 4aa, respectively) of P. chrysogenum, P. vulpinum, and P. expansum fused to the RGS-His-tag were cloned into PCR cloning vector pDrive (Qiagen, Hild ...
... RGSHHHHHH-tag in case of the prp8-ir. The PCR amplification products containing the prp8 intein coding sequence with flanking extein sequences (5aa and 4aa, respectively) of P. chrysogenum, P. vulpinum, and P. expansum fused to the RGS-His-tag were cloned into PCR cloning vector pDrive (Qiagen, Hild ...
i ANALYSIS OF THE ENU-3 PROTEIN FAMILY IN NERVOUS
... incomplete. Additionally, the interface between these cues and their receptors as well as how these interactions allow cells to make decisions regarding their correct migration pathways during development, are challenging aspects of the nervous system that are still under investigation. It is import ...
... incomplete. Additionally, the interface between these cues and their receptors as well as how these interactions allow cells to make decisions regarding their correct migration pathways during development, are challenging aspects of the nervous system that are still under investigation. It is import ...
Denaturation of hemoglobin in the presence of tannic acid
... and is found in numerous plants and foods. It generally can be extracted from grape seeds. TA is known as an effective antioxidant, antimicrobial/antiviral agent, which binds to blood proteins [5–8]. The binding of a ligand to HSA and HHb can stabilize or destabilize the protein structure. Fluoresce ...
... and is found in numerous plants and foods. It generally can be extracted from grape seeds. TA is known as an effective antioxidant, antimicrobial/antiviral agent, which binds to blood proteins [5–8]. The binding of a ligand to HSA and HHb can stabilize or destabilize the protein structure. Fluoresce ...
Green fluorescent protein
The green fluorescent protein (GFP) is a protein composed of 238 amino acid residues (26.9 kDa) that exhibits bright green fluorescence when exposed to light in the blue to ultraviolet range. Although many other marine organisms have similar green fluorescent proteins, GFP traditionally refers to the protein first isolated from the jellyfish Aequorea victoria. The GFP from A. victoria has a major excitation peak at a wavelength of 395 nm and a minor one at 475 nm. Its emission peak is at 509 nm, which is in the lower green portion of the visible spectrum. The fluorescence quantum yield (QY) of GFP is 0.79. The GFP from the sea pansy (Renilla reniformis) has a single major excitation peak at 498 nm.In cell and molecular biology, the GFP gene is frequently used as a reporter of expression. In modified forms it has been used to make biosensors, and many animals have been created that express GFP as a proof-of-concept that a gene can be expressed throughout a given organism. The GFP gene can be introduced into organisms and maintained in their genome through breeding, injection with a viral vector, or cell transformation. To date, the GFP gene has been introduced and expressed in many Bacteria, Yeast and other Fungi, fish (such as zebrafish), plant, fly, and mammalian cells, including human. Martin Chalfie, Osamu Shimomura, and Roger Y. Tsien were awarded the 2008 Nobel Prize in Chemistry on 10 October 2008 for their discovery and development of the green fluorescent protein.