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... Understanding the transcriptional regulation networks that control the gene expression programs in a living cell is one of fundamental questions in molecular biology. Using single perturbations allows us to identify the key players in those networks. To better understand the structure, we can use do ...
... Understanding the transcriptional regulation networks that control the gene expression programs in a living cell is one of fundamental questions in molecular biology. Using single perturbations allows us to identify the key players in those networks. To better understand the structure, we can use do ...
Microtubule Associated Protein 1b (MAP1B) Is a Marker of the
... and a high postnatal mortality. Although it was noted that they appeared malnourished and dehydrated, the cause of their growth reduction as well as of their high mortality has not been established [16]. We speculated that MAP1B is necessary for kidney glomerular development and function, thereby ex ...
... and a high postnatal mortality. Although it was noted that they appeared malnourished and dehydrated, the cause of their growth reduction as well as of their high mortality has not been established [16]. We speculated that MAP1B is necessary for kidney glomerular development and function, thereby ex ...
Insightful directed evolution of Escherichia coli quorum sensing
... lsr operon promoters using error-prone PCR (ePCR) and selected for promoters that provided E. coli with higher tetracycline resistance over the native promoter when placed upstream of the tet(C) gene. Among the fourteen clones identified, we found several mutations in the binding sites of QS repress ...
... lsr operon promoters using error-prone PCR (ePCR) and selected for promoters that provided E. coli with higher tetracycline resistance over the native promoter when placed upstream of the tet(C) gene. Among the fourteen clones identified, we found several mutations in the binding sites of QS repress ...
Localised axial progenitor cell populations in the avian tail bud are
... Production of GFP-expressing transgenic chickens for fate-mapping experiments We previously reported that the CMV-IE enhancer/promoter drove limited reporter gene expression in transgenic chickens (McGrew et al., 2004). By contrast, high-level, widespread expression has been achieved using a compoun ...
... Production of GFP-expressing transgenic chickens for fate-mapping experiments We previously reported that the CMV-IE enhancer/promoter drove limited reporter gene expression in transgenic chickens (McGrew et al., 2004). By contrast, high-level, widespread expression has been achieved using a compoun ...
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... Production of GFP-expressing transgenic chickens for fate-mapping experiments We previously reported that the CMV-IE enhancer/promoter drove limited reporter gene expression in transgenic chickens (McGrew et al., 2004). By contrast, high-level, widespread expression has been achieved using a compoun ...
... Production of GFP-expressing transgenic chickens for fate-mapping experiments We previously reported that the CMV-IE enhancer/promoter drove limited reporter gene expression in transgenic chickens (McGrew et al., 2004). By contrast, high-level, widespread expression has been achieved using a compoun ...
The Golgi-Localized Arabidopsis Endomembrane
... EMP12 Is a Golgi-Localized Protein with Multiple TMDs All 12 isoforms of Arabidopsis EMP proteins (termed EMP1 to EMP12 in this study; see Supplemental Figure 1 online) share high similarity at the amino acid level and in predicted topology. All At EMPs are predicted to have a large lumenal N-termin ...
... EMP12 Is a Golgi-Localized Protein with Multiple TMDs All 12 isoforms of Arabidopsis EMP proteins (termed EMP1 to EMP12 in this study; see Supplemental Figure 1 online) share high similarity at the amino acid level and in predicted topology. All At EMPs are predicted to have a large lumenal N-termin ...
A Cytoplasm to Vacuole Targeting Pathway in P. pastoris
... conditions of PpApe1‑CFP in the absence of P. pastoris orthologs of proteins involved in all autophagy‑related pathways (PpAtg5, PpAtg8, PpAtg9, and PpVps15), or in the absence of two proteins (PpAtg11 and PpVac8) required only for the Cvt pathway in S. cerevisiae (Fig. 2A and B). Whereas in wild‑t ...
... conditions of PpApe1‑CFP in the absence of P. pastoris orthologs of proteins involved in all autophagy‑related pathways (PpAtg5, PpAtg8, PpAtg9, and PpVps15), or in the absence of two proteins (PpAtg11 and PpVac8) required only for the Cvt pathway in S. cerevisiae (Fig. 2A and B). Whereas in wild‑t ...
Environmentally Sensitive Fluorescent Sensors Based on Synthetic Peptides Linköping University Post Print
... synthesis. Barbara Imperiali’s group has developed the functionalized amino acids DANA [33] and 4-DAPA (4-N,N-dimethylaminophtalimidoalanine, [35]), and used them to probe the phosphorylation-dependent binding of an octapeptide to a 14-3-3 protein. The 14-3-3 proteins are essential intermediates in ...
... synthesis. Barbara Imperiali’s group has developed the functionalized amino acids DANA [33] and 4-DAPA (4-N,N-dimethylaminophtalimidoalanine, [35]), and used them to probe the phosphorylation-dependent binding of an octapeptide to a 14-3-3 protein. The 14-3-3 proteins are essential intermediates in ...
SLR-2 and JMJC-1 regulate an evolutionarily conserved stress
... elegans has emerged as an excellent system for the study of stress networks and the underlying principles that govern their regulation. One recurring theme is that the majority of stress-response pathways exhibit some degree of pleiotropy in their activating conditions. For example, HSF-1/HSF1 is ac ...
... elegans has emerged as an excellent system for the study of stress networks and the underlying principles that govern their regulation. One recurring theme is that the majority of stress-response pathways exhibit some degree of pleiotropy in their activating conditions. For example, HSF-1/HSF1 is ac ...
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... 2008; Olmedo-Verd et al., 2011). Membrane anchoring was particularly well demonstrated for the Valyl-tRNA synthetase (ValRS) of Anabaena sp. PCC 7120 (hereafter Anabaena), which harbors an idiosyncratic domain, termed CAAD, with a putative C-terminal coiled-coil and two transmembrane helices, that m ...
... 2008; Olmedo-Verd et al., 2011). Membrane anchoring was particularly well demonstrated for the Valyl-tRNA synthetase (ValRS) of Anabaena sp. PCC 7120 (hereafter Anabaena), which harbors an idiosyncratic domain, termed CAAD, with a putative C-terminal coiled-coil and two transmembrane helices, that m ...
Transgenic line for the identification of cholinergic release sites in
... antibody staining. Because of the small diameter of the neuronal processes, reliable localization of the antibody staining within the individual neurites is beyond the resolution threshold of traditional confocal microscopy. Therefore, the markers that localize to the presynaptic regions and are not ...
... antibody staining. Because of the small diameter of the neuronal processes, reliable localization of the antibody staining within the individual neurites is beyond the resolution threshold of traditional confocal microscopy. Therefore, the markers that localize to the presynaptic regions and are not ...
Power tools for gene expression and clonal analysis in
... 18 °C but does not repress GAL4 at 29 °C or higher temperatures. Alternatively, one can use recombinases to temporally control the expression of GAL80, as we will discuss below. This threecomponent system (GAL4-UAS, transgene and GAL80) is the basis of two powerful technologies: MARCM (discussed bel ...
... 18 °C but does not repress GAL4 at 29 °C or higher temperatures. Alternatively, one can use recombinases to temporally control the expression of GAL80, as we will discuss below. This threecomponent system (GAL4-UAS, transgene and GAL80) is the basis of two powerful technologies: MARCM (discussed bel ...
Dominant-lethal alpha-tubulin mutants defective in microtubule depolymerization in yeast.
... relative to those of the tubulin crystal structure, due to an additional amino acid at position 44). D252 is conserved in nearly all tubulins and in bacterial FtsZ (which is structurally nearly identical), whereas E255 is specific to ␣-tubulins (Nogales et al., 1998a). The TUB1-828 phenotype is domi ...
... relative to those of the tubulin crystal structure, due to an additional amino acid at position 44). D252 is conserved in nearly all tubulins and in bacterial FtsZ (which is structurally nearly identical), whereas E255 is specific to ␣-tubulins (Nogales et al., 1998a). The TUB1-828 phenotype is domi ...
Extended PDF
... was seen in adjacent gfp cells (Figure 1Ca). However, after 30 min, gfp cells lying in proximity to gfp+ cells acquired a weak fluorescence signal (Figure 1Cb). The fluorescence intensity of gfp cells increased over time in a manner inversely proportional to their distance from the gfp+ cells, i. ...
... was seen in adjacent gfp cells (Figure 1Ca). However, after 30 min, gfp cells lying in proximity to gfp+ cells acquired a weak fluorescence signal (Figure 1Cb). The fluorescence intensity of gfp cells increased over time in a manner inversely proportional to their distance from the gfp+ cells, i. ...
University of Groningen A new set of small, extrachromosomal
... minimal fragment containing only the origin of replication and the G5 gene can still support extrachromosomal replication (Kiyosawa et al., 1995), but plasmids containing this minimal fragment are rapidly lost from the population when selective pressure is removed, indicating that replication and pa ...
... minimal fragment containing only the origin of replication and the G5 gene can still support extrachromosomal replication (Kiyosawa et al., 1995), but plasmids containing this minimal fragment are rapidly lost from the population when selective pressure is removed, indicating that replication and pa ...
De Novo Pyrimidine Nucleotide Synthesis Mainly
... cells showed uptake of [14C]-uracil in a time-dependent manner. Uracil uptake was linear for at least 5 min and then accounted for 3.75 mmol mg21 protein, whereas noninduced cells imported only 0.52 mmol mg21 protein (Figure 3A). The apparent affinity of PLUTO for uracil was 16.4 mM (Figure 3B). A d ...
... cells showed uptake of [14C]-uracil in a time-dependent manner. Uracil uptake was linear for at least 5 min and then accounted for 3.75 mmol mg21 protein, whereas noninduced cells imported only 0.52 mmol mg21 protein (Figure 3A). The apparent affinity of PLUTO for uracil was 16.4 mM (Figure 3B). A d ...
De Novo Pyrimidine Nucleotide Synthesis Mainly
... (i.e., nucleobases or nucleosides) are imported into plastids. A corresponding uracil transport activity could be measured in intact plastids isolated from cauliflower (Brassica oleracea) buds. PLUTO (for plastidic nucleobase transporter) was identified as a member of the Nucleobase:Cation-Symporter ...
... (i.e., nucleobases or nucleosides) are imported into plastids. A corresponding uracil transport activity could be measured in intact plastids isolated from cauliflower (Brassica oleracea) buds. PLUTO (for plastidic nucleobase transporter) was identified as a member of the Nucleobase:Cation-Symporter ...
An Enhanced System for Unnatural Amino Acid
... properties of proteins with exquisite control over structure. This approach has a number of potential advantages over other “chemical mutagenesis” methods such as the use of chemically aminoacylated tRNAs, solid-phase peptide synthesis, native chemical ligation, or intein-based semisynthetic methods ...
... properties of proteins with exquisite control over structure. This approach has a number of potential advantages over other “chemical mutagenesis” methods such as the use of chemically aminoacylated tRNAs, solid-phase peptide synthesis, native chemical ligation, or intein-based semisynthetic methods ...
In Vivo Selection Yields AAV-B1 Capsid for Central
... comprised of 60 subunits of VP1, VP2, and VP3 capsid proteins in a ratio of ~1:1:10, and an encapsidated single-stranded DNA viral genome. In addition to protecting the genome, the capsid mediates interactions with cell surface receptors and postentry intracellular trafficking and as such, is the ma ...
... comprised of 60 subunits of VP1, VP2, and VP3 capsid proteins in a ratio of ~1:1:10, and an encapsidated single-stranded DNA viral genome. In addition to protecting the genome, the capsid mediates interactions with cell surface receptors and postentry intracellular trafficking and as such, is the ma ...
2.3.1. Vector construction - Trace: Tennessee Research and
... Biological interactions occur on multiple length scales, ranging from molecular to population wide interactions. This work describes the study of two specific areas of biological interactions in microbial systems: intracellular protein-protein interactions and cell-to-cell interactions. The implemen ...
... Biological interactions occur on multiple length scales, ranging from molecular to population wide interactions. This work describes the study of two specific areas of biological interactions in microbial systems: intracellular protein-protein interactions and cell-to-cell interactions. The implemen ...
Characterization of a novel phosphatidylinositol 3
... Mowiol 40-88 (Aldrich, Gillingham, Dorset, U.K.)\1,4diazadicyclo[2.2.2]octane (‘ DABCO ’) and allowed to dry before examination on a Zeiss LSM 410 confocal laser scanning microscope with excitation wavelengths of 488 nm (enhanced yellow fluorescent protein) and 543 nm (Texas Red). ...
... Mowiol 40-88 (Aldrich, Gillingham, Dorset, U.K.)\1,4diazadicyclo[2.2.2]octane (‘ DABCO ’) and allowed to dry before examination on a Zeiss LSM 410 confocal laser scanning microscope with excitation wavelengths of 488 nm (enhanced yellow fluorescent protein) and 543 nm (Texas Red). ...
Blimp1 regulates development of the posterior forelimb, caudal
... (see Fig. S2 in the supplementary material). Thus mutant forelimbs entirely lacked the ulna, and either two (7/15) or three (8/15) of the posterior digits, but were otherwise indistinguishable from the wild type. The signalling centres controlling outgrowth and patterning of the early limb bud are w ...
... (see Fig. S2 in the supplementary material). Thus mutant forelimbs entirely lacked the ulna, and either two (7/15) or three (8/15) of the posterior digits, but were otherwise indistinguishable from the wild type. The signalling centres controlling outgrowth and patterning of the early limb bud are w ...
... also identified a separate Wnt pathway, involving the Frz receptor LIN-17 and the Wnt ligands LIN-44 and CWN-1, that preserves VA inputs by blocking CEH-12 expression in anterior VAs. Our results have uncovered a key role for the UNC-4 transcription factor in modulating the relative strengths of Wnt ...
Redox States of Plastids and Mitochondria
... to specifically alter the redox state of chloroplasts or mitochondria and then determined their impact on PD-mediated intercellular transport. Paraquat (also called methyl viologen dichloride) is a herbicide that primarily affects plants by acting as a terminal oxidant for PSI and by direct oxidatio ...
... to specifically alter the redox state of chloroplasts or mitochondria and then determined their impact on PD-mediated intercellular transport. Paraquat (also called methyl viologen dichloride) is a herbicide that primarily affects plants by acting as a terminal oxidant for PSI and by direct oxidatio ...
Endocytosis of cigarette-smoke condensate by rabbit alveolar
... were incubated with the CSC. When the phagocytic cell population (monocytes) were examined in the flow cytofluorometer an almost threefold increase in fluorescence was observed at 37°C compared to 4°C. This was in contrast to the non-phagocytic cells (lymphocytes) which only showed a slightly enhanc ...
... were incubated with the CSC. When the phagocytic cell population (monocytes) were examined in the flow cytofluorometer an almost threefold increase in fluorescence was observed at 37°C compared to 4°C. This was in contrast to the non-phagocytic cells (lymphocytes) which only showed a slightly enhanc ...
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.