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  JoVE Developmental Biology


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Genetics: The branch of science concerned with the means and consequences of transmission and generation of the components of biological inheritance. (Stedman, 26th ed)
 JoVE Immunology and Infection

Forward Genetics Screens Using Macrophages to Identify Toxoplasma gondii Genes Important for Resistance to IFN-γ-Dependent Cell Autonomous Immunity

1Department of Microbiology and Immunology, New York Medical College

JoVE 52556

Forward genetics is a powerful approach to identify genes in intracellular pathogens important for resistance to cell autonomous immunity. The current approach uses innate immune cells, specifically macrophages, to identify novel Toxoplasma gondii genes important for immune evasion.

 JoVE Behavior

Automated, Quantitative Cognitive/Behavioral Screening of Mice: For Genetics, Pharmacology, Animal Cognition and Undergraduate Instruction

1Department of Psychology, Rutgers University, 2Department of Psychology, Koç University, 3Center for Neural Science, New York University, 4Department of Mathematics & Computer Science, Fairfield University

JoVE 51047

Fully automated system for measuring physiologically meaningful properties of the mechanisms mediating spatial localization, temporal localization, duration, rate and probability estimation, risk assessment, impulsivity, and the accuracy and precision of memory, in order to assess the effects of genetic and pharmacological manipulations on foundational mechanisms of cognition in mice.

 JoVE Immunology and Infection

VIGS-Mediated Forward Genetics Screening for Identification of Genes Involved in Nonhost Resistance

1Plant Biology Division, The Samuel Roberts Noble Foundation

JoVE 51033

Virus-induced gene silencing is an useful tool for identifying genes involved in nonhost resistance of plants. We demonstrate the use of bacterial pathogens expressing GFPuv in identifying gene silenced plants susceptible to nonhost pathogens. This approach is easy, fast and facilitates large scale screening and similar protocol can be applied to studying various other plant-microbe interactions.

 JoVE Immunology and Infection

Reverse Genetics Mediated Recovery of Infectious Murine Norovirus

1Section of Virology, Imperial College London

JoVE 4145

Noroviruses are a major cause of gastroenteritis yet molecular techniques for their characterisation are still relatively new. Here we report two different reverse genetics approaches for the efficient recovery of murine norovirus (MNV), the only member of this genus which can be propagated in cell culture.

 JoVE Immunology and Infection

Using Reverse Genetics to Manipulate the NSs Gene of the Rift Valley Fever Virus MP-12 Strain to Improve Vaccine Safety and Efficacy

1Department of Pathology, University of Texas Medical Branch

JoVE 3400

The reverse genetics system for the Rift Valley fever virus MP-12 vaccine strain is a useful tool for creating additional MP-12 mutants with increased attenuation and immunogenicity. We describe the protocol to generate and characterize NSs mutant strains.

 Science Education: Essentials of Developmental Biology

An Introduction to Developmental Genetics

JoVE Science Education

Development is the complex process through which a single-celled embryo transforms into a multicellular organism. Developmental processes are guided by information encoded in an organism\'s DNA, and geneticists are trying to understand how this information leads to a fully formed organism.

This video reviews seminal research in the field of developmental biology, including the identification of specific genes that control various embryonic processes. An introduction to the major questions asked by developmental geneticists, and the prominent methods used to answer them, is also provided. Finally, several applications of these prominent methods are discussed, in order to show specific experiments currently being performed in this field.

 Science Education: Essentials of Genetics

An Overview of Genetics and Disease

JoVE Science Education

Many human diseases are associated with mutations or variations in genetic sequences. Some of these genetic variants are heritable, passed down from generation to generation, while others arise sporadically during an organism’s life and cause diseases such as cancer. Researchers are trying to identify and characterize these genetic alterations in the hopes of improving diagnosis and therapeutic options for patients.In this video, we will examine the history of genetic disease research, and explore key questions asked by medical geneticists. Various tools used to identify the genetic basis of diseases are then discussed, including genotyping techniques and genome-wide association studies (GWAS). Finally, several current examples of medical genetics research are presented.

 JoVE Biology

A Manual Small Molecule Screen Approaching High-throughput Using Zebrafish Embryos

1Department of Biological Sciences, University of Notre Dame

JoVE 52063

The zebrafish is an excellent experimental organism to study vertebrate developmental processes and model human disease. Here, we describe a protocol on how to perform a manual high-throughput chemical screen in zebrafish embryos with a whole-mount in situ hybridization (WISH) read-out.

 JoVE Biology

Aseptic Laboratory Techniques: Plating Methods

1Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles

JoVE 3064

When working with media and reagents used to culture microorganisms, aseptic technique must be practiced to ensure contamination is minimized. A variety of plating methods are routinely used to isolate, propagate, or enumerate bacteria and phage, all of which incorporate procedures that maintain the sterility of experimental materials.

 JoVE Biology

Assessing Differences in Sperm Competitive Ability in Drosophila

1Department of Ecology and Evolutionary Biology, University of California, Irvine

JoVE 50547

Differential sperm competitive ability among Drosophila males with distinct genotypes can be ascertained through double-mating experiments. Each of these experiments involves one of the males of interest and a reference male. Readily identifiable markers in the progeny allow inference of the fraction of individuals fathered by each male.

 JoVE Biology

Culturing Caenorhabditis elegans in Axenic Liquid Media and Creation of Transgenic Worms by Microparticle Bombardment

1Department of Animal and Avian Sciences, University of Maryland, 2Department of Cell Biology and Molecular Genetics, University of Maryland

JoVE 51796

C. elegans is usually grown on solid agar plates or in liquid cultures seeded with E. coli. To prevent bacterial byproducts from confounding toxicological and nutritional studies, we utilized an axenic liquid medium, CeHR, to grow and synchronize a large number of worms for a range of downstream applications.

 JoVE Biology

Purification of Transcripts and Metabolites from Drosophila Heads

1Department of Neurology, McKnight Brain Institute, University of Florida, 2Department of Entomology and Nematology, University of Florida, 3Genetics Institute, Department of Molecular Genetics and Microbiology, University of Florida, 4McKnight Brain Institute, Department of Neuroscience, Genetics Institute, Center for Translational Research on Neurodegenerative Diseases, and Center for Movement Disorders and Neurorestoration, University of Florida

JoVE 50245

We describe here the procedures for the extraction and purification of mRNA and metabolites from Drosophila heads. We are applying these techniques to better understand the cellular perturbations underlying neuronal degeneration. These methodologies can be easily scaled and adapted for other "omic" projects.

 JoVE Immunology and Infection

Generation of Recombinant Arenavirus for Vaccine Development in FDA-Approved Vero Cells

1Department of Microbiology and Immunology, University of Rochester School of Medicine and Dentistry, 2Departments of Immunology and Microbial Sciences, The Scripps Research Institute

JoVE 50662

Rescue of recombinant arenaviruses from cloned cDNAs, an approach referred to as reverse genetics, allows researchers to investigate the role of specific viral gene products, as well as the contribution of their different specific domains and residues, to many different aspects of the biology of arenavirus. Likewise, reverse genetics techniques in FDA-approved cell lines (Vero) for vaccine development provides novel possibilities for the generation of effective and safe vaccines to combat human pathogenic arenaviruses.

 JoVE Biology

Imaging Centrosomes in Fly Testes

1Department of Biological Sciences, University of Toledo

JoVE 50938

Imaging of centrosomal proteins during Drosophila spermatogenesis is a powerful method to identify new proteins critical for centrosome biology as well as to elucidate the particular function of known players in this process.

 JoVE Biology

Growth-based Determination and Biochemical Confirmation of Genetic Requirements for Protein Degradation in Saccharomyces cerevisiae

1Department of Biology, Ball State University, 2Division of Nephrology, Cincinnati Children's Hospital

JoVE 52428

This article describes a yeast growth-based assay for the determination of genetic requirements for protein degradation. It also demonstrates a method for rapid extraction of yeast proteins, suitable for western blotting to biochemically confirm degradation requirements. These techniques can be adapted to monitor degradation of a variety of proteins.

 JoVE Biology

BEST: Barcode Enabled Sequencing of Tetrads

1Pacific Northwest Diabetes Research Institute

JoVE 51401

Barcode Enabled Sequencing of Tetrads (BEST) replaces the manual processes of isolating, disrupting and spacing tetrads. BEST isolates tetrads by fluorescence-activated cell sorting onto agar plates, separates the spores by agitation with glass beads, and determines which randomly arrayed colonies were derived from the same original tetrad using molecular barcodes.

 JoVE Biology

Generation of High Quality Chromatin Immunoprecipitation DNA Template for High-throughput Sequencing (ChIP-seq)

1Division of Human Genetics, Children's Hospital of Philadelphia Research Institute, 2Department of Pediatrics, Perelman School of Medicine, University of Pennsylvania

JoVE 50286

The combination of chromatin immunoprecipitation and ultra-high-throughput sequencing (ChIP-seq) can identify and map protein-DNA interactions in a given tissue or cell line. Outlined is how to generate a high quality ChIP template for subsequent sequencing, using experience with the transcription factor TCF7L2 as an example.

 JoVE Neuroscience

An Inexpensive, Scalable Behavioral Assay for Measuring Ethanol Sedation Sensitivity and Rapid Tolerance in Drosophila

1Department of Human and Molecular Genetics, Virginia Commonwealth University, 2VCU Alcohol Research Center, Virginia Commonwealth University

JoVE 52676

Straightforward assays for measuring ethanol sensitivity and rapid tolerance in Drosophila facilitate the use of this model organism for investigating these important ethanol-related behaviors. Here, a relatively simple, scalable assay for measuring ethanol sensitivity and rapid tolerance in flies is described.

 JoVE Neuroscience

A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

1School of Life Sciences, Arizona State University

JoVE 51057

The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species.

 JoVE Neuroscience

Visualizing Neuroblast Cytokinesis During C. elegans Embryogenesis

1Department of Biology, Concordia University

JoVE 51188

This protocol describes how to image dividing cells within a tissue in Caenorhabditis elegans embryos. While several protocols describe how to image cell division in the early embryo, this protocol describes how to image cell division within a developing tissue during mid late embryogenesis.

 JoVE Developmental Biology

Comprehensive Assessment of Germline Chemical Toxicity Using the Nematode Caenorhabditis elegans

1Institute for Society and Genetics, University of California, Los Angeles, 2Environmental Health Sciences, University of California, Los Angeles, 3California Nanosystems Institute, Department of Molecular and Medical Pharmacology, University of California, Los Angeles

JoVE 52445

We describe the detailed steps of a high-throughput chemical assay in the nematode Caenorhabditis elegans used to assess germline toxicity. In this assay, disruption of germline function following chemical exposure is monitored using a fluorescent reporter specific to aneuploid embryos.

 JoVE Immunology and Infection

Intranasal Administration of Recombinant Influenza Vaccines in Chimeric Mouse Models to Study Mucosal Immunity

1Laboratory of Emerging Viruses, Heinrich Pette Institute, Leibniz Institute for Experimental Virology

JoVE 52803

There is an overall lack of knowledge about how vaccines work. Here we propose the combined use of reverse genetics and bone marrow chimeric mice to gain insight into the early host immune responses to vaccines with a special focus on dendritic cells and T cell immunity.

 JoVE Biology

Biochemical and High Throughput Microscopic Assessment of Fat Mass in Caenorhabditis Elegans

1Center for Human Genetic Research and Department of Medicine, Massachusetts General Hospital and Harvard Medical School, 2Department of Earth, Atmospheric, and Planetary Sciences, Massachusetts Institute of Technology

JoVE 50180

We present robust biochemical and microscopic methods for studying Caenorhabditis elegans lipid stores. A rapid, simple, fixing-staining procedure for fluorescent lipid droplet imaging leverages the spectral properties of the lipophilic dye Nile red. We then present biochemical measurement of triglycerides and phospholipids using solid phase extraction and gas chromatography-mass spectrometry.

 JoVE Biology

Recombineering Homologous Recombination Constructs in Drosophila

1Department of Molecular Biology, University of Texas Southwestern Medical Center at Dallas, 2Department of Physiology, University of Texas Southwestern Medical Center at Dallas, 3Green Center for Systems Biology, University of Texas Southwestern Medical Center at Dallas

JoVE 50346

Homologous recombination techniques greatly advance Drosophila genetics by enabling the creation of molecularly precise mutations. The recent adoption of recombineering allows one to manipulate large pieces of DNA and transform them into Drosophila6. The methods presented here combine these techniques to rapidly generate large homologous recombination vectors.

 JoVE Immunology and Infection

Modeling The Lifecycle Of Ebola Virus Under Biosafety Level 2 Conditions With Virus-like Particles Containing Tetracistronic Minigenomes

1Laboratory of Virology, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 2Research Technology Branch, Division of Intramural Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health

JoVE 52381

Work with infectious Ebola viruses is restricted to biosafety level 4 laboratories. Tetracistronic minigenome-containing replication and transcription-competent virus like particles (trVLPs) represent a lifecycle modeling system that allows us to safely model multiple infectious cycles under biosafety level 2 conditions, relying exclusively on Ebola virus components.

 JoVE Biology

Reporter-based Growth Assay for Systematic Analysis of Protein Degradation

1Department of Biological Chemistry, The Hebrew University of Jerusalem

JoVE 52021

Here we describe a robust biological assay for quantifying the relative rate of proteolysis by the ubiquitin-proteasome system. The assay readout is yeast growth rate in liquid culture, which is dependent on the cellular levels of a reporter protein comprising a degradation signal fused to an essential metabolic marker.

 JoVE Biology

Combining Magnetic Sorting of Mother Cells and Fluctuation Tests to Analyze Genome Instability During Mitotic Cell Aging in Saccharomyces cerevisiae

1Department of Biological Sciences, Rensselaer Polytechnic Institute

JoVE 51850

Mutation rates in young Saccharomyces cerevisiae cells measured through fluctuation tests are used to predict mutation frequencies for mother cells of different replicative ages. Magnetic sorting and flow cytometry are then used to measure actual mutation frequencies and age of mother cells to identify any deviations from predicted mutation frequencies.

 JoVE Biology

Measuring the Effects of Bacteria on C. Elegans Behavior Using an Egg Retention Assay

1Department of Biological and Allied Health Sciences, Fairleigh Dickinson University

JoVE 51203

An egg-in-worm (EIW) assay is a useful method to quantify egg-laying behavior. Alterations in egg laying can be a behavioral response of the model organism Caenorhabditis elegans to potentially harmful environmental substances such as those produced by pathogenic bacteria.

 JoVE Neuroscience

In Vivo Imaging of Dauer-specific Neuronal Remodeling in C. elegans

1Department of Crop Sciences, University of Illinois Urbana-Champaign

JoVE 51834

Following exposure to specific environmental stressors, the nematode Caenorhabditis elegans undergoes extensive phenotypic plasticity to enter into a stress-resistant ‘dauer’ juvenile stage. We present methods for the controlled induction and imaging of neuroplasticity during dauer.

 JoVE Biology

Mouse Genome Engineering Using Designer Nucleases

1Institute of Laboratory Animal Science, University of Zurich, 2Department of Genetics, Cell Biology & Development and Center for Genome Engineering, University of Minnesota

JoVE 50930

Designer nucleases such as zinc finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs) can be used to modify the genome of mouse preimplantation embryos by triggering both the nonhomologous end joining (NHEJ) and homologous recombination (HR) pathways. These advances enable the rapid generation of mice with precise genetic modifications.

 JoVE Biology

A Video Protocol of Retroviral Infection in Primary Intestinal Organoid Culture

1Department of Genetics, University of Cambridge, 2Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge

JoVE 51765

This protocol explains primary Lgr5-positve organoid culture and the subsequent performance of retroviral transduction. This enables Cre-inducible overexpression or knockdown of the delivered transgene and allows functional studies to be carried out in the novel in vitro organotypic model system.

 JoVE Biology

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

1Department of Biological Sciences, Center for Applied Chemical Biology, Youngstown State University

JoVE 51934

Enterobacter sp. YSU grows in glucose minimal salts medium. Auxotrophs are generated by transforming it with a transposome which randomly inserts itself into the host genome. Mutants are found by replica plating from complex medium to minimal medium. Interrupted genes are identified by gene rescue and sequencing.

 JoVE Biology

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization

1Department of Pathology, New York University School of Medicine, 2New York University Center for Health Informatics and Bioinformatics, 3NYU Cancer Institute, 4Department of Pathology and Yale Cancer Center, Yale University School of Medicine

JoVE 50087

Here we describe a protocol for simultaneous detection of histone modifications by immunofluorescence and DNA sequences by DNA FISH followed by 3D microscopy and analyses (3D immuno-DNA FISH).

 JoVE Biology

Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

1Harvard Medical School, 2Division of Hematology/Oncology, Boston Children's Hospital, 3Department of Pediatric Oncology, Dana-Farber Cancer Institute, 4Howard Hughes Medical Institute

JoVE 52118

CRISPR/Cas9 is a robust system to produce disruption of genes and genetic elements. Here we describe a protocol for the efficient creation of genomic deletions in mammalian cell lines using CRISPR/Cas9.

 JoVE Immunology and Infection

Rescue of Recombinant Newcastle Disease Virus from cDNA

1Department of Microbiology, Icahn School of Medicine at Mount Sinai, 2Global Health and Emerging Pathogens Institute, Icahn School of Medicine at Mount Sinai, 3Department of Medicine, Icahn School of Medicine at Mount Sinai, 4Department of Microbiology and Immunology, School of Medicine and Dentistry, University of Rochester

JoVE 50830

Newcastle disease virus (NDV) has been extensively studied in the last few years in order to develop new vectors for vaccination and therapy, among others. These studies have been possible due to techniques to rescue recombinant virus from cDNA, such as those we describe here.

 JoVE Biology

Enhanced Reduced Representation Bisulfite Sequencing for Assessment of DNA Methylation at Base Pair Resolution

1Department of Medicine, Weill Cornell Medical College, 2Institute for Computational Biomedicine, Weill Cornell Medical College, 3Department of Physiology and Biophysics, Weill Cornell Medical College, 4Department of Pathology, University of Michigan

JoVE 52246

Enhanced Reduced Representation Bisulfite Sequencing is a method for the preparation of sequencing libraries for DNA methylation analysis based on restriction enzyme digestion combined with cytosine bisulfite conversion. This protocol requires 50 ng of starting material and yields base pair resolution data at GC-rich genomic regions.

 JoVE Biology

Measuring Oxidative Stress Resistance of Caenorhabditis elegans in 96-well Microtiter Plates

1Goodman Cancer Research Center, McGill University, 2Department of Biochemistry, McGill University

JoVE 52746

C. elegans is an attractive model organism to study signal transduction pathways involved in oxidative stress resistance. Here we provide a protocol to measure oxidative stress resistance of C. elegans animals in liquid phase, using several oxidizing agents in 96 well plates.

 JoVE Biology

Production of Haploid Zebrafish Embryos by In Vitro Fertilization

1Department of Biological Sciences, University of Notre Dame

JoVE 51708

The zebrafish is a powerful model system for developmental biology and human disease research due to their genetic similarity with higher vertebrates. This protocol describes a methodology to create haploid zebrafish embryos that can be utilized for forward screen strategies to identify recessive mutations in genes essential for early embryogenesis.

 JoVE Medicine

Collection and Extraction of Saliva DNA for Next Generation Sequencing

1Battelle Center for Mathematical Medicine, The Research Institute at Nationwide Children's Hospital, 2The Interdisciplinary Graduate Program in Biophysics, The Ohio State University, 3Department of Pediatrics, The Ohio State University

JoVE 51697

DNA extraction from saliva can provide a readily available source of high molecular weight DNA, with little to no degradation/fragmentation. This protocol provides optimized parameters for saliva collection/storage and DNA extraction to be of sufficient quality and quantity for downstream DNA assays with high quality requirements.

 JoVE Biology

Examination of Drosophila Larval Tracheal Terminal Cells by Light Microscopy

1Department of Human Genetics, University of Utah

JoVE 50496

Here, we present a method for light microscopy analysis of tracheal terminal cells in Drosophila larvae. This method allows for quick examination of branch and lumen morphology in whole animals and would be useful for analysis of individual mutants or screens for mutations affecting terminal cell development.

 JoVE Environment

Methods for Performing Crosses in Setaria viridis, a New Model System for the Grasses

1Donald Danforth Plant Science Center, 2Boyce Thompson Institute

JoVE 50527

We have developed a methodology for performing crosses in Setaria viridis (S. viridis). The method involves pruning the panicle prior to a hot water treatment to kill viable pollen. Crosses are performed following a well-controlled growth regime and typically result in the recovery of 1 to 7 cross-pollinated seed/s per panicle.

 JoVE Biology

Methods to Assess Subcellular Compartments of Muscle in C. elegans

1MRC/ARUK Centre for Musculoskeletal Ageing Research, University of Nottingham

JoVE 52043

Skeletal muscle is essential for locomotion and is the bodies’ main protein store. Muscle health measurements within C. elegans are described. Prospective changes to muscle structure and function are assessed using localized GFP and cationic dyes.

 JoVE Biology

Using Whole Mount in situ Hybridization to Link Molecular and Organismal Biology

1Department of Biology, Syracuse University, 2Department of Science Teaching, Syracuse University

JoVE 2533

Whole mount in situ hybridization (WISH) was used in an upper level undergraduate Comparative Vertebrate Biology course in addition to vertebrate dissections. This gave students the opportunity to study gene expression patterns as well as gross anatomy, linking the study of molecular and organismal biology within one course.

 JoVE Biology

Stable Isotopic Profiling of Intermediary Metabolic Flux in Developing and Adult Stage Caenorhabditis elegans

1Department of Pediatrics, The Children's Hospital of Philadelphia, 2Department of Pediatrics, University of Pennsylvania

JoVE 2288

Stable isotopic profiling by gas chromatography mass spectrometric analysis of intermediary metabolic flux is described in the nematode, Caenorhabditis elegans. Methods are detailed for assessing isotopic enrichment in carbon dioxide, organic acids, and amino acids following isotope exposure either during development on agar plates or during adulthood in liquid culture.

 JoVE Biology

Dietary Supplementation of Polyunsaturated Fatty Acids in Caenorhabditis elegans

1School of Molecular Biosciences, Washington State University, 2Center for Reproductive Biology, Washington State University

JoVE 50879

Caenorhabditis elegans is a useful model to explore the functions of polyunsaturated fatty acids in development and physiology. This protocol describes an efficient method of supplementing the C. elegans diet with polyunsaturated fatty acids.

 JoVE Neuroscience

Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

1Department of Pathology and Laboratory Medicine, University of North Carolina School of Medicine, 2Lineberger Comprehensive Cancer Center, University of North Carolina School of Medicine, 3Division of Neuropathology, Department of Pathology and Laboratory Medicine, University of North Carolina School of Medicine, 4Curriculum in Genetics and Molecular Biology, University of North Carolina School of Medicine, 5Biological and Biomedical Sciences Program, University of North Carolina School of Medicine, 6Department of Radiation Oncology, Emory University School of Medicine, 7Department of Neurology, Neurosciences Center, University of North Carolina School of Medicine

JoVE 51763

Phenotypically wild-type astrocytes and neural stem cells harvested from mice engineered with floxed, conditional oncogenic alleles and transformed via viral Cre-mediated recombination can be used to model astrocytoma pathogenesis in vitro and in vivo by orthotopic injection of transformed cells into brains of syngeneic, immune-competent littermates.

 JoVE Biology

RNAi Screening to Identify Postembryonic Phenotypes in C. elegans

1Department of Molecular and Cellular Medicine, Texas A&M University System Health Science Center

JoVE 3442

We describe a sensitized method to identify postembryonic regulators of protein expression and localization in C. elegans using an RNAi-based genomic screen and an integrated transgene that expresses a functional, fluorescently tagged protein.

 JoVE Biology

Optimization and Utilization of Agrobacterium-mediated Transient Protein Production in Nicotiana

1Fraunhofer USA Center for Molecular Biotechnology

JoVE 51204

Transient protein production in Nicotiana plants based on vacuum infiltration with Agrobacteria carrying launch vectors (Tobacco mosaic virus-based) is a rapid and economic approach to produce vaccine antigens and therapeutic proteins. We simplified the procedure and improved target accumulation by optimizing conditions of bacteria cultivation, selecting host species, and co-introducing RNA silencing suppressors.

 JoVE Neuroscience

Denver Papillae Protocol for Objective Analysis of Fungiform Papillae

1Health Sciences Department, Denver Museum of Nature & Science, 2Department of Mathematical and Statistical Sciences, University of Colorado Denver

JoVE 52860

Here we present a protocol to measure fungiform papilla density from digital photographs. This method builds prioritization and objective characteristic metrics into the original descriptive work on fungiform papillae by Miller & Reedy (1990).

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