• Volume 12, 2022
  • Volume 11, 2021
  • Volume 10, 2020
  • Volume 9, 2019
  • Volume 8, 2018
  • Volume 7, 2017
  • Volume 6, 2016
  • Volume 5, 2015
  • Volume 4, 2014
  • Volume 3, 2013
  • Volume 2, 2012
  • Volume 1, 2011

Biological Engineering

Site-specific Incorporation of Phosphoserine into Recombinant Proteins in Escherichia coli Authors:  Phillip Zhu, Ryan A. Mehl and Richard B. Cooley, date: 11/05/2022, view: 231, Q&A: 0

This protocol describes the recombinant expression of proteins in E. coli containing phosphoserine (pSer) installed at positions guided by TAG codons. The E. coli strains that can be used here are engineered with a ∆serB genomic knockout to produce pSer internally at high levels, so no exogenously added pSer is required, and the addition of pSer

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A Fluorescence-based Approach Utilizing Self-labeling Enzyme Tags to Determine Protein Orientation in Large Unilamellar Vesicles Authors:  Laura Charlotte Paweletz, Sarina Veit and Thomas Günther Pomorski, date: 11/05/2022, view: 248, Q&A: 0

Reconstitution of membrane proteins into large unilamellar vesicles is an essential approach for their functional analysis under chemically defined conditions. The orientation of the protein in the liposomal membrane after reconstitution depends on many parameters, and its assessment is important prior to functional measurements. Common approaches

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Biophysics

In situ cryo-FIB/SEM Specimen Preparation Using the Waffle Method Authors:  Oleg Klykov, Daija Bobe, Mohammadreza Paraan, Jake D. Johnston, Clinton S. Potter, Bridget Carragher, Mykhailo Kopylov and Alex J. Noble, date: 11/05/2022, view: 634, Q&A: 0

Cryo-focused ion beam (FIB) milling of vitrified specimens is emerging as a powerful method for in situ specimen preparation. It allows for the preservation of native and near-native conditions in cells, and can reveal the molecular structure of protein complexes when combined with cryo-electron tomography (cryo-ET) and sub-tomogram averaging.

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Binding Affinity Measurements Between DNA Aptamers and their Virus Targets Using ELONA and MST Authors:  Gregory T. Pawel, Yuan Ma, Yuting Wu, Yi Lu and Ana Sol Peinetti, date: 11/05/2022, view: 286, Q&A: 0

Aptamers have been selected with strong affinity and high selectivity for a wide range of targets, as recently highlighted by the development of aptamer-based sensors that can differentiate infectious from non-infectious viruses, including human adenovirus and SARS-CoV-2. Accurate determination of the binding affinity between the DNA aptamers and

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Cancer Biology

OxiDIP-Seq for Genome-wide Mapping of Damaged DNA Containing 8-Oxo-2'-Deoxyguanosine Authors:  Francesca Gorini, Giovanni Scala, Susanna Ambrosio, Barbara Majello and Stefano Amente, date: 11/05/2022, view: 357, Q&A: 0

8-oxo-7,8-dihydro-2′-deoxyguanosine (8-oxodG) is considered to be a premutagenic DNA lesion generated by 2'-deoxyguanosine (dG) oxidation due to reactive oxygen species (ROS). In recent years, the 8-oxodG distribution in human, mouse, and yeast genomes has been underlined using various next-generation sequencing (NGS)–based strategies.

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An Unbiased CRISPR-Cas9 Screening Method for the Identification of Positive and Negative Regulatory Proteins of Cell Adhesion Authors:  Yvonne J. Thus, Martin F. M. de Rooij, Roderick L. Beijersbergen and Marcel Spaargaren, date: 11/05/2022, view: 222, Q&A: 0

Mature B-cell lymphomas are highly dependent upon the protective lymphoid organ microenvironment for their growth and survival. Targeting integrin-mediated homing and retention of the malignant B cells in the lymphoid organs, using the Bruton’s tyrosine kinase (BTK) inhibitor ibrutinib, is a highly efficacious FDA-approved therapy for

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Immunology

Murine Double Hit Model for Neonatal Cardiopulmonary Diseases: Bronchopulmonary Dysplasia (BPD) and Pulmonary Hypertension Associated with BPD Authors:  Steven P. Garrick, Philip J. Berger, Marcel F. Nold and Claudia A. Nold-Petry, date: 11/05/2022, view: 276, Q&A: 0

Bronchopulmonary dysplasia (BPD) and pulmonary hypertension associated with BPD (BPD-PH) are of multifactorial origin and share common risk factors. Most murine models of BPD expose newborn pups to only one of these risk factors—more commonly postnatal hyperoxia—thereby mimicking the vital increased fraction of inspired oxygen (FiO2)

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Mechanobiology

Motion-capture Analysis of Mice Using a Video Recorded on an iPhone Camera Authors:  Ryo Nakamichi and Hiroshi Asahara, date: 11/05/2022, view: 420, Q&A: 0

When focusing on quick movements in the analysis of animal behavior, a high-speed camera can be used as a powerful tool. There are many options for high-speed cameras to record movement. In recent years, the quality and sophistication of videos captured on cell phones have evolved so much that the iPhone’s slow-motion video system can

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Microbiology

In vitro Di-ubiquitin Formation Assay and E3 Cooperation Assay Authors:  Rebecca J. Burge, Katie H. Jameson, Anthony J. Wilkinson and Jeremy C. Mottram, date: 11/05/2022, view: 221, Q&A: 0

Ubiquitination is a post-translational modification conserved across eukaryotic species. It contributes to a variety of regulatory pathways, including proteasomal degradation, DNA repair, and cellular differentiation. The ubiquitination of substrate proteins typically requires three ubiquitination enzymes: a ubiquitin-activating E1, a

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Plant Science

X-ray Crystallography: Seeding Technique with Cytochrome P450 Reductase Authors:  Bixia Zhang, Jacob A. Lewis, Rishi Hazra and ChulHee Kang, date: 11/05/2022, view: 237, Q&A: 0

Cytochrome P450 reductase (CPR) is a multi-domain protein that acts as a redox partner of cytochrome P450s. The CPR contains a flavin adenine dinucleotide (FAD)–binding domain, a flavin mononucleotide (FMN)-binding domain, and a connecting domain. To achieve catalytic events, the FMN-binding domain needs to move relative to the FAD-binding

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