JoVE Logo

Sign In

University of Colorado at Boulder

5 ARTICLES PUBLISHED IN JoVE

image

Biology

Transmembrane Domain Oligomerization Propensity determined by ToxR Assay
Catherine Joce 1, Alyssa Wiener 1, Hang Yin 1
1Department of Chemistry and Biochemistry, University of Colorado at Boulder

An efficient procedure to assess the oligomerization propensity of single-pass transmembrane domains (TMDs) is described. Chimeric proteins consisting of the TMD fused to ToxR are expressed in an E. coli reporter strain. TMD-induced oligomerization causes dimerization of ToxR, activation of transcription and production of the reporter protein, -galactosidase.

image

Bioengineering

Constant Pressure-controlled Extrusion Method for the Preparation of Nano-sized Lipid Vesicles
Leslie A. Morton *1,2, Jonel P. Saludes *1,2, Hang Yin 1,2
1Department of Chemistry & Biochemistry, University of Colorado Boulder, 2Biofrontiers Institute, University of Colorado Boulder

This protocol describes an extrusion method for preparing lipid vesicles of sub-micron sizes with a high degree of homogeneity. This method uses a pressure-controlled system with controlled nitrogen flow rates for liposome preparation. The lipid preparation1,2, liposome extrusion, and size characterization will be presented herein.

image

Bioengineering

In Vitro Model of Physiological and Pathological Blood Flow with Application to Investigations of Vascular Cell Remodeling
Winston Elliott 1, Devon Scott-Drechsel 2, Wei Tan 1
1Department of Mechanical Engineering, University of Colorado at Boulder, 2Product Development, Covidien, Ltd.

This protocol replicates physiological or pathological blood flow in vitro to aid in determining cell response in disease pathologies. By introducing a pressure damping chamber downstream of a blood pump, blood flow across the vasculature can be recapitulated and imposed on a monolayer of vascular endothelium or a mimetic co-culture.

image

Genetics

A Novel Saturation Mutagenesis Approach: Single Step Characterization of Regulatory Protein Binding Sites in RNA Using Phosphorothioates
Ravinder Singh 1
1Department of Molecular, Cellular and Developmental Biology, University of Colorado at Boulder

Proteins that bind specific RNA sequences play critical roles in gene expression. Detailed characterization of these binding sites is crucial for our understanding of gene regulation. Here, a single-step approach for saturation mutagenesis of protein-binding sites in RNA is described. This approach is relevant for all protein-binding sites in RNA.

image

Genetics

Exploring Sequence Space to Identify Binding Sites for Regulatory RNA-Binding Proteins
Ravinder Singh 1
1Department of Molecular, Cellular and Developmental Biology, University of Colorado at Boulder

Sequence specificity is critical for gene regulation. Regulatory proteins that recognize specific sequences are important for gene regulation. Defining functional binding sites for such proteins is a challenging biological problem. An iterative approach for identification of a binding site for an RNA-binding protein is described here and is applicable to all RNA-binding proteins.

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved