227 Result(s)
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Protocol
Production of Double-Stranded RNA Using the Prokaryotic Promoter-Mediated Bidirectional Transcription
Large-scale and cost-less production of double-stranded RNA (dsRNA) is the basis for the widespread application of dsRNA in agriculture. Bidirectional transcription of target sequence in RNase III-deficient Esche...
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Protocol
Application of dsRNA in the Pine Wood Nematode, Bursaphelenchus xylophilus
The pine wood nematode Bursaphelenchus xylophilus is one of the most destructive invasive species worldwide, causing the wilting and eventual death of pine trees. Despite recognition of their economic and environ...
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Protocol
Genetic Code Expansion in Pseudomonas putida KT2440
Emerging microorganism Pseudomonas putida KT2440 is utilized for the synthesis of biobased chemicals from renewable feedstocks and for bioremediation. However, the methods for analyzing, engineering, and regulati...
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Protocol
Identification of Mycoviruses by dsRNA Extraction
Mycoviruses exist in all major groups of fungi. With the continuous development of science and technology, the methods of studying viruses are constantly updated, and progressively mycoviruses have been discov...
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Protocol
An Overview of High-Throughput Crop Phenoty**: Platform, Image Analysis, Data Mining, and Data Management
In this chapter, we explore the application of high-throughput crop phenoty** facilities for phenotype data acquisition and the extraction of significant information from the collected data through image pro...
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Protocol
Genetically Encoded Noncanonical Amino Acids in Proteins to Investigate Lysine Benzoylation
Posttranslational modifications (PTMs) of lysine residues are major regulators of gene expression, protein–protein interactions, and protein localization and degradation. Histone lysine benzoylation is a recen...
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Protocol
Visualization of Exo- and Endocytosis Membrane Dynamics with Super-Resolution STED Microscopy
Recent advances in stimulated emission depletion (STED) microscopy offer an unparalleled avenue to study membrane dynamics of exo- and endocytosis, such as fusion pore opening, pore expansion, constriction, an...
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Protocol
Surface Modification of Liposomes Using Folic Acid
Liposomes are usually defined as spherically shaped microscopic vesicles that consist of one or more phospholipid bilayer membranes. They are widely used in drug delivery due to their biocompatibility, biodegr...
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Protocol
High-Throughput and In-Depth Proteomic Profiling of 5 μL Plasma and Serum Using TMTpro 16-Plex
High-throughput and in-depth proteomic analysis of plasma and serum samples remains challenging due to the presence of multiple high-abundance proteins. Here, we provide a detailed protocol for proteomic analy...
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Protocol
Reconstitution of Vesicle Budding from the TGN and Immunoisolation of Vesicles Enriched with a Specific Cargo Client
The trans-Golgi network (TGN) is an important cargo sorting station in the secretory pathway. To reveal insights into protein sorting at the TGN, it is important to develop an assay to quantify the efficiency of ...
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Protocol
qPCR Genoty** of Polyploid Species
A simple and cost-effective method for genoty** polyploid plants using quantitative PCR (qPCR) is described in this chapter. There is no additional operation, only simultaneous amplification of alleles and r...
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Protocol
Preparation and Fractionation of Heterogeneous Aβ42 Oligomers with Different Aggregation Properties
Deposition of amyloid-β (Aβ) aggregates in the form of amyloid plaques is a central feature of Alzheimer’s disease. The end products of Aβ aggregation are amyloid fibrils. Soluble Aβ aggregates called oligomer...
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Protocol
Lysosome Imaging Based on Fluorescent Carbon Dots
Lysosomes play key roles in different cellular processes such as autophagy, phagocytosis, and apoptosis. Lysosomal dysfunction is related to many diseases. Fluorescence lysosome staining strategy is valuable f...
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Protocol
Genetic Code Expansion in Mammalian Cells Through Quadruplet Codon Decoding
Genetic code expansion enables the site-specific incorporation of noncanonical amino acids (ncAAs) into proteins both in vitro and in vivo. In addition to a widely applied nonsense suppression strategy, the us...
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Protocol
Site-Specific Incorporation of Sulfotyrosine into Proteins in Mammalian Cells
Protein (PTS) plays a crucial role in numerous extracellular protein-protein interactions. It is involved in diverse physiological processes and the development of human diseases, including AIDS and cancer. ...
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Protocol
Engineering of SH2 Domains for the Recognition of Protein Tyrosine O-Sulfation Sites
Protein engineering has brought advances to industrial processes, biomaterials, nanotechnology, biosensors, and biomedical applications. This chapter will focus on the engineering of Src Homology 2 domains (SH...
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Protocol
Predicting Pseudouridine Sites with Porpoise
is a ubiquitous modification and plays a crucial role in many biological processes. However, it remains a challenging task to identify sites using expensive and time-consuming experimental research. To th...
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Protocol
Introduction of Genome Editing Reagents and Genoty** of Derived Edited Alleles in Soybean (Glycine max (L.) Merr.)
Cas9-based genome editing is a powerful genetic tool for loci specifically targeted for genome modification. This chapter describes up-to-date protocols using Cas9-based genome editing technology, including ve...
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Protocol
Imaging of Cortical Microtubules in Plants Under Salt Stress
The microtubule (MT) network is a highly dynamic subcellular structure playing an important role in the growth and development of plants, and it is able to respond to biotic and abiotic environmental signals. ...
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Protocol
Assessing SIRT7 Activity In Vivo and In Vitro in Response to DNA Damage
The class III histone deacetylase (HDACs) also known as sirtuins (SIRTs 1–7) are ubiquitously expressed, but SIRT7 mainly resides as nucleolar protein. In this chapter a couple of methods are described that ar...
