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    Transgene Recombineering in Bacterial Artificial Chromosomes

    Bacterial Artificial Chromosome (BAC) libraries are a valuable research resource. Any one of the clones in these libraries can carry hundreds of thousands of base pairs of genetic information. Often the entire...

    Michael G. Zeidler, Thomas L. Saunders in Microinjection (2019)

  2. No Access

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    Chemical Synthesis of an Asymmetric Mimic of the Nitrogenase Active Site

    The synthetic inorganic chemistry of metal–sulfur (M-S, M = metals) clusters has played an important, complementary role to the biochemical analyses of nitrogenase toward a better understanding of the enzyme a...

    Kazuki Tanifuji, Yasuhiro Ohki in Metalloproteins (2019)

  3. No Access

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    Interface-Based Structural Prediction of Novel Host-Pathogen Interactions

    About 20% of the cancer incidences worldwide have been estimated to be associated with infections. However, the molecular mechanisms of exactly how they contribute to host tumorigenesis are still unknown. To e...

    Emine Guven-Maiorov, Chung-Jung Tsai in Computational Methods in Protein Evolution (2019)

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    In Silico-Directed Evolution Using CADEE

    Recent years have seen an explosion of interest in both sequence- and structure-based approaches toward in silico-directed evolution. We recently developed a novel computational toolkit, CADEE, which facilitat...

    Beat Anton Amrein, Ashish Runthala in Computational Methods in Protein Evolution (2019)

  5. No Access

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    Rare Variant Quantitation Using Droplet Digital PCR

    Droplet digital PCR (ddPCR) allows precise quantitation of individual copies of DNA. Using allele-specific fluorescent oligonucleotide probes, ddPCR can enumerate both mutant and wild-type alleles enabling hig...

    Brian Parkin in Chronic Lymphocytic Leukemia (2019)

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    Genome Sequencing and Analysis Methods in Chronic Lymphocytic Leukemia

    The genomic sequencing of chronic lymphocytic leukemia (CLL) samples has provided exciting new venues for the understanding and treatment of this prevalent disease. This feat is possible thanks to high-through...

    Víctor Quesada, Miguel Araujo-Voces, José G. Pérez-Silva in Chronic Lymphocytic Leukemia (2019)

  7. No Access

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    Expression, Purification, and Activity Analysis of Chlorophyllide Oxidoreductase and Ni2+-Sirohydrochlorin a,c-Diamide Reductase

    Nitrogenase-like enzymes play a vital role in the reduction of the conjugated ring systems of diverse tetrapyrrole molecules. The biosynthesis of all bacteriochlorophylls involves the two-electron reduction of...

    Jürgen Moser, Jan Jasper, José Vazquez Ramos, Sven T. Sowa in Metalloproteins (2019)

  8. No Access

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    Proteomic Detection of Carbohydrate-Active Enzymes (CAZymes) in Microbial Secretomes

    Secretomes from microorganisms growing on biomass contain carbohydrate-active enzymes (CAZymes) of potential biotechnological interest. By analyzing such secretomes, we may discover key enzymes involved in deg...

    Tina R. Tuveng, Vincent G. H. Eijsink, Magnus Ø. Arntzen in Functional Proteomics (2019)

  9. No Access

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    Protocols for the Molecular Evolutionary Analysis of Membrane Protein Gene Duplicates

    Gene duplication is an important process in the evolution of gene content in eukaryotic genomes. Understanding when gene duplicates contribute new molecular functions to genomes through molecular adaptation is...

    Laurel R. Yohe, Liang Liu, Liliana M. Dávalos in Computational Methods in Protein Evolution (2019)

  10. No Access

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    Measuring Human Lipid Metabolism Using Deuterium Labeling: In Vivo and In Vitro Protocols

    Stable isotopes are powerful tools for tracing the metabolic fate of molecules in the human body. In this chapter, we focus on the use of deuterium (2H), a stable isotope of hydrogen, in the study of human lipid ...

    Katherine E. Pinnick, Pippa J. Gunn, Leanne Hodson in Metabolic Signaling (2019)

  11. No Access

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    A Cell-Free Content Mixing Assay for SNARE-Mediated Multivesicular Body-Vacuole Membrane Fusion

    Endocytosis is a fundamental process underlying diverse eukaryotic physiology. The terminal stage of this process is membrane fusion between the perimeter membrane of a late endosome filled with intraluminal v...

    Mahmoud Abdul Karim, Dieter Ronny Samyn, Christopher Leonard Brett in SNAREs (2019)

  12. No Access

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    Quantifying Intramolecular Protein Conformational Dynamics Under Lipid Interaction Using smFRET and FCCS

    Fӧrster-type resonance energy transfer (FRET) with fluorescence cross-correlation spectroscopy (FCCS) is a powerful combination for observing intramolecular conformational dynamics on the micro- to millisecond...

    Pei Li, Yawei Dai, Markus Seeger, Yan-Wen Tan in SNAREs (2019)

  13. No Access

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    Differential Scanning Fluorimetry and Hydrogen Deuterium Exchange Mass Spectrometry to Monitor the Conformational Dynamics of NBD1 in Cystic Fibrosis

    Cystic fibrosis (CF) is one of the most common, lethal autosomal recessive diseases in Caucasians with a life expectancy of 37–47 years. The CF transmembrane conductance regulator (CFTR) is a plasma membrane i...

    Naoto Soya, Ariel Roldan, Gergely L. Lukacs in Protein Misfolding Diseases (2019)

  14. No Access

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    Quantification of Protein Aggregates Using Bimolecular Fluorescence Complementation

    Neuronal accumulations of the presynaptic protein α-synuclein represent a hallmark of Parkinson’s disease (PD). Multiple system atrophy and dementia with Lewy bodies are other neurodegenerative diseases in wh...

    Vibha Prasad, Aaron Voigt in Protein Misfolding Diseases (2019)

  15. No Access

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    Generating Genetically Engineered Mice Using a Spermatogonial Stem Cell-Mediated Method

    Mouse spermatogonial stem cells (SSCs) can be grown in culture for long periods. Cultured SSCs, also called germline stem (GS) cells, maintain themselves by self-renewing proliferation while retaining the abil...

    Takuya Sato, Takehiko Ogawa in Microinjection (2019)

  16. No Access

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    High-Throughput Sequencing of the T-Cell Receptor Beta Chain Gene Repertoire in Chronic Lymphocytic Leukemia

    High-throughput, next-generation sequencing (NGS) offers a unique opportunity for in-depth characterization of adaptive immune receptor repertoires. Nevertheless, limitations and pitfalls exist in every step o...

    E. Vlachonikola, A. Vardi, K. Stamatopoulos in Chronic Lymphocytic Leukemia (2019)

  17. No Access

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    Generation of Conditional Knockout Mice by Sequential Insertion of Two loxP Sites In Cis Using CRISPR/Cas9 and Single-Stranded DNA Oligonucleotides

    Conditional knockout (cKO) mice are extremely valuable for biomedical research because they enable detailed analyses of gene functions in a tissue- or temporally-specific fashion. The conventional method for g...

    Ye Liu, Yubin Du, Wen Xie, Fan Zhang, Douglas Forrest, Chengyu Liu in Microinjection (2019)

  18. No Access

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    Autophagy in Adipocyte Differentiation

    Adipose tissue regulates metabolic homeostasis by acting as an endocrine organ and energy reservoir. Adipose tissue development and functional maintenance are dependent on adipocyte differentiation, in which a...

    Zhipeng Tao, Longhua Liu, Louise D. Zheng in Autophagy in Differentiation and Tissue Ma… (2019)

  19. No Access

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    Functional Proteomic Analysis to Characterize Signaling Crosstalk

    The biological activities of a cell are determined by its response to external stimuli. The signals are transduced from either intracellular or extracellular milieu through networks of multi-protein complexes ...

    Sneha M. Pinto, Yashwanth Subbannayya, T. S. Keshava Prasad in Functional Proteomics (2019)

  20. No Access

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    Coevolutionary Signals and Structure-Based Models for the Prediction of Protein Native Conformations

    The analysis of coevolutionary signals from families of evolutionarily related sequences is a recent conceptual framework that provides valuable information about unique intramolecular interactions and, theref...

    Ricardo Nascimento dos Santos, Xianli Jiang in Computational Methods in Protein Evolution (2019)

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