*  Molecular Characterization of Binding of Substrates and Inhibitors to DT-Diaphorase: Combined Approach Involving Site-Directed...
In summary, by evaluating results obtained from a combined approach involving site-directed mutagenesis of DT-diaphorase, ... Combined Approach Involving Site-Directed Mutagenesis, Inhibitor-Binding Analysis, and Computer Modeling. Shiuan Chen, Kebin Wu ... Combined Approach Involving Site-Directed Mutagenesis, Inhibitor-Binding Analysis, and Computer Modeling. Shiuan Chen, Kebin Wu ... Combined Approach Involving Site-Directed Mutagenesis, Inhibitor-Binding Analysis, and Computer Modeling. Shiuan Chen, Kebin Wu ...
  http://molpharm.aspetjournals.org/content/56/2/272
*  A novel role for DYX1C1, a chaperone protein for both Hsp70 and Hsp90, in breast cancer | SpringerLink
Hsp70 mutant constructs were assembled using a PCR-based, site-directed mutagenesis approach. The plasmid F2 (pGADT7-GFP-EEVD) ... GST pull-down, a yeast two-hybrid system, RT-PCR, site-directed mutagenesis approach. ... site (GGCCATTACGGCC); 3′-Sfi-I (B) site (GGCCGAGGCGGCC)] separated by a sufficient number of basepairs to allow efficient ... The Sfi-I fragment of the product was ligated into the Sfi-I (A) and Sfi-I (B) sites of vectors, pGBDT7 and pGEX-4T-1. The ...
  https://link.springer.com/article/10.1007%2Fs00432-009-0568-6
*  Directed Mutagenesis of Nicotinic Receptors to Investigate Receptor Function | IntechOpen
Directed Mutagenesis of Nicotinic Receptors to Investigate Receptor Function , IntechOpen, Published on: 2013-02-05. Authors: ... Similarly, a site-directed mutagenesis approach was taken to generate 36 toxin mutants. The objective was to identify the ... Use of directed mutagenesis in nicotinic acetylcholine receptor research. Site-directed mutagenesis is a powerful tool to ... In summary, a combined approach of site-directed mutagenesis experiments, molecular modelling and docking studies was needed to ...
  https://www.intechopen.com/books/genetic-manipulation-of-dna-and-protein-examples-from-current-research/directed-mutagenesis-of-nicotinic-receptors-to-investigate-receptor-function/
*  KAKEN - Research Projects | Protein engineering for conferring a biological function on ovalbumin (KAKENHI-PROJECT-15380229)
We have done site-directed mutagenesis approach to confer serpin function on ovalbumin. We found by X-ray crystallography that ... For further mutagenesis approach, we created a different ovalbumin mutant R339T/A352R in which the P1-P1' site is accessible ... 12,000,000 (Direct Cost : ¥12,000,000). Fiscal Year 2004 : ¥5,800,000 (Direct Cost : ¥5,800,000). Fiscal Year 2003 : ¥6,200,000 ... We therefore did further mutagenesis to accelerate the loop insertion rate on the basis of the data of crystal structure. ...
  https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15380229/
*  Identification of residues/sequences in the human riboflavin transporter-2 that is important for function and cell biology |...
... site-directed mutagenesis, and truncation approaches. The human-derived brain U87 cells were used as a model in these ... site-directed mutagenesis, and truncation approach in the human-derived brain U87 cell model. ... Site-directed mutagenesis and generation of the N- and C-terminal truncated hRFVT-2 construct. Studies described in the ... Point mutations in the open reading frame (ORF) of hRFVT-2 were introduced using the Quick Change site-directed mutagenesis kit ...
  https://nutritionandmetabolism.biomedcentral.com/articles/10.1186/s12986-015-0008-3
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... Although this approach provides higher sensitivity, it generally needs prior sample pre-fractionation 7 and pre-digestion with ... The direct analysis by LC-MS without trypsin digestion showed that human saliva indigenously contains many peptide fragments ... Transcranial direct current stimulation (tDCS) is a neuromodulation technique that has been increasingly used over the past ...
  https://www.jove.com/visualize/abstract/22615919/clinical-severity-thalassaemiahb-e-disease-is-associated-with
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... Previous approaches to study sensitization are generally cumbersome involving continuous cell culture maintenance as well as a ... PIP3 serve as binding sites for proteins with pleckstrin Homology (PH) domains, thus recruiting these proteins to the membrane ... Studying the role of a protein in post-meiotic spermatogenesis using mutagenesis or other genetic tools is often impeded by ...
  https://www.jove.com/visualize/abstract/23028418/fsh-modulates-pkai-gpr3-activities-mouse-oocyte-coc-gap-junctional
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach results in transfection of over 90% cultured RPTC. Here, we present methods for assessing the role of PKC-ε in: 1 ... Using this approach, 50-60% of mice develop moderate AKI 24 hr after injury but 90-100% of mice survive. To induce more severe ... However, this approach leads to high background of non-interacting proteins making it difficult to interpret co-localization ...
  https://www.jove.com/visualize/abstract/22768207/cd98-increases-renal-epithelial-cell-proliferation-by-activating-mapks
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach can be used to isolate nuclei from a wide variety of Drosophila embryonic and larval cell types using specific ... To validate this approach, we dissected equivalent areas of the disc from the posterior (labelled by GFP expression), and the ... The approach described in this protocol enables consistent isolation of nuclei from specific cell types in the Drosophila ...
  https://www.jove.com/visualize/abstract/23056495/anion-sensitive-fluorophore-identifies-drosophila-swell-activated
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach has been employed to analyze effects of viral mutations on HIV-1 capsid stability4, 5, 6. It should also be ... This approach, termed Tandem Affinity Purification (TAP), was initially used in yeast2,3 but more recently has been adapted to ... The approach involves isolation of cores by sedimentation of concentrated virions through a layer of detergent and into a ...
  https://www.jove.com/visualize/abstract/21042582/the-initial-step-human-immunodeficiency-virus-type-1-gagpropol
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... The first two experimental approaches monitor yeast growth on plates, the third approach monitors the growth of liquid yeast ... Transcranial direct current stimulation, tDCS, High-definition transcranial direct current stimulation, HD-tDCS, Electrical ... Our approach is to use hippocampal slice cultures as an in vitro model that closely reflects their in vivo counterparts with ...
  https://www.jove.com/visualize/abstract/19488402/ikkalpha-ikkbeta-regulation-dna-damage-induced-cleavage
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach can be used to isolate nuclei from a wide variety of Drosophila embryonic and larval cell types using specific ... The module approach generates uniform, scalable and vascularized tissues. The modules can be made of collagen as well as other ... The approach described in this protocol enables consistent isolation of nuclei from specific cell types in the Drosophila ...
  https://www.jove.com/visualize/abstract/21779388/a-conformation-sensitive-monoclonal-antibody-against-a2-domain-von
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... The interaction was direct and did not involve intermediate molecules. By mass spectrometry, we established the identity of the ... In parallel to the electrophysiological experiments, a biotinylation approach was used to monitor the appearance of γENaC ... or to compare the transport capacity and kinetics of site-specifically mutated transporters with high precision. Furthermore, ...
  https://www.jove.com/visualize/abstract/19238202/localisation-function-endocannabinoid-system-human
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... Transgenic mice carrying site-specific genome modifications (knockout, knock-in) are of vital importance for dissecting complex ... of designer nuclease target sites within a gene of interest followed by construction of custom DNA-binding domains to direct ... RNA sequence elements in the 3' UTR of the mRNA, the Vg1 localization element (VLE) are required and sufficient to direct ...
  https://www.jove.com/visualize/abstract/24194906/resveratrol-protects-mouse-oocytes-from-methylglyoxal-induced
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... site-directed mutagenesis, G protein-coupled receptor, targeted photocrosslinking, bioorthogonal labeling, targeted epitope ... Although chemical mutagenesis using N-ethyl-N-nitrosourea (ENU) or ethyl methanesulfonate (EMS) has been used for decades in ... Here, we report methods for using type I and type II Δku80Δhxgprt strains to advance gene targeting approaches in T. gondii. We ...
  https://www.jove.com/visualize/abstract/24781326/structural-basis-for-immunomodulatory-function-cysteine-protease
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach can be used to isolate nuclei from a wide variety of Drosophila embryonic and larval cell types using specific ... Our approach is based on metabolic labeling of newly transcribed RNA with 4-thiouridine (thus also referred to as 4sU-tagging) ... To validate this approach, we dissected equivalent areas of the disc from the posterior (labelled by GFP expression), and the ...
  https://www.jove.com/visualize/abstract/19834616/probe-level-analysis-expression-microarrays-characterizes-isoform
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... FAQs and video tutorials are available at the SCOPE web site which also includes a "Sample Search" button that allows the user ... In this article, we utilize a web version of SCOPE2 to examine genes that are involved in telomere maintenance. SCOPE has been ... Advantages of the miRNA assay approach used in this study is that it enables a fast screening of mature miRNA regulations in ...
  https://www.jove.com/visualize/abstract/22348050/genome-wide-identification-immune-response-analysis-serine-protease
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach improves upon existing methods in that many current protocols use submerged monolayer or Transwell cultures of ... A direct laryngoscopy was attempted with no visualization of the larynx. Oxygen saturation was difficult to maintain, with ... Additionally, the nose is the entry point to the respiratory system and therefore one of the first sites to be exposed to any ...
  https://www.jove.com/visualize/abstract/23741370/human-neutrophil-elastase-degrades-splunc1-impairs-airway-epithelial
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... whereas the direct injection method addresses the virulence of whole-cell inocula. The utility of the direct injection method ... An approach is described that utilizes the host innate immune response, in this case macrophages, in a forward genetic screen ... These approaches allow for the rapid screening of mutants to determine factors important in L. pneumophila virulence, ...
  https://www.jove.com/visualize/abstract/25803442/shrimp-serine-proteinase-homologues-pmmassph-1-2-play-role-activation
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach results in transfection of over 90% cultured RPTC. Here, we present methods for assessing the role of PKC-ε in: 1 ... This approach can be adapted to any other cellular body and granule protein component, and performed with transgenic animals, ... However, this approach leads to high background of non-interacting proteins making it difficult to interpret co-localization ...
  https://www.jove.com/visualize/abstract/23342057/ack1-activation-and-regulation-by-allostery
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... The experimental approach using laboratory-based sediment incubations to quantify internal P load is a direct method, making it ... Our approach is to use hippocampal slice cultures as an in vitro model that closely reflects their in vivo counterparts with ... Our approach is based on metabolic labeling of newly transcribed RNA with 4-thiouridine (thus also referred to as 4sU-tagging) ...
  https://www.jove.com/visualize/abstract/24676354/reference-gene-selection-for-gene-expression-analysis-oocytes
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... This approach results in transfection of over 90% cultured RPTC. Here, we present methods for assessing the role of PKC-ε in: 1 ... To demonstrate this approach, herein BOMUS and AIM MRI are used to map unilateral mechanical stimulation of the vibrissae in ... A New Approach for the Comparative Analysis of Multiprotein Complexes Based on 15N Metabolic Labeling and Quantitative Mass ...
  https://www.jove.com/visualize/abstract/21448286/the-serine-814-trpc6-is-phosphorylated-under-unstimulated
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... Our approach is to use hippocampal slice cultures as an in vitro model that closely reflects their in vivo counterparts with ... By using random mutagenesis we identified RSP_1090 to be required for full activation of the RpoE response under (1)O2 stress, ... The direct functionalization of C-H bonds is an important and long standing goal in organic chemistry. Such transformations can ...
  https://www.jove.com/visualize/abstract/24223961/degs-rsep-homologous-proteases-are-involved-singlet-oxygen-dependent
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... Although this approach provides higher sensitivity, it generally needs prior sample pre-fractionation 7 and pre-digestion with ... To approach this problem, we have introduced peptide array based substrate specificity analyses of PKMTs. Peptide arrays are ... The direct analysis by LC-MS without trypsin digestion showed that human saliva indigenously contains many peptide fragments ...
  https://www.jove.com/visualize/abstract/19492082/a-sequence-structure-based-method-to-predict-putative-substrates
*  JoVE | Peer Reviewed Scientific Video Journal - Methods and Protocols
In combination with site-directed mutagenesis, this approach provides a powerful tool to identify functionally relevant ... The primary advantage of utilizing directed evolution instead of more rational-based approaches for molecular engineering ... Toward this end, directed evolution has been shown to be a useful tool for altering the thermal activity of various molecules ... Directed evolution is defined as a method to harness natural selection in order to engineer proteins to acquire particular ...
  https://www.jove.com/visualize/abstract/26087013/clonal-evolution-blast-crisis-correlate-with-enhanced-proteolytic