INTRODUCTION TO GENETIC CIRCUITS AND RECOMBINANT PROTEIN EXPRESSION
Genetic circuits are a set of interconnected genes and regulatory elements that work together to control cellular processes. In the context of optimizing recombinant protein expression, genetic circuits can be designed to enhance the production of desired proteins within host…
PROTEIN STEREOCHEMICAL AND SECONDARY STRUCTURE ANALYSIS
1) Protein Stereochemical Analysis Protein stereochemistry refers to the three-dimensional arrangement of atoms in a protein molecule. It plays a crucial role in determining the structure, stability, and function of proteins. Understanding protein stereochemistry is essential for various applications in…
PROTEIN DOMAINS AS STRUCTURAL, FUNCTIONAL, AND EVOLUTIONARY UNITS
Proteins are essential macromolecules that play a crucial role in various biological processes. They are composed of one or more structural and functional units called protein domains. Protein domains are distinct regions within a protein sequence that fold into a…
GENERATION OF SELECTIVE PROTEIN BINDING SEQUENCES IN GLYCOSAMINOGLYCANS
Glycosaminoglycans (GAGs) are linear polysaccharides that are an essential component of the extracellular matrix and play crucial roles in various biological processes, including cell signaling, tissue development, and inflammation. One area of significant interest is the generation and identification of…
THE CRITICAL FUNCTION OF CO-TRANSLATIONAL GLYCOSYLATION ON PROTEIN FOLDING AND SECRETION
Co-translational glycosylation is a crucial post-translational modification process that plays a significant role in protein folding and secretion. This process involves the addition of oligosaccharide chains to nascent polypeptides as they are being synthesized by ribosomes. The glycosylation of proteins…
A COMPREHENSIVE LOOK AT PROTEIN FUSION TAGS
Protein fusion tags are commonly used in molecular biology to aid in the purification, detection, and expression of recombinant proteins. These tags are typically short amino acid sequences that are genetically fused to the protein of interest. They can facilitate…
GENERAL STEPS INVOLVED IN DEVELOPING PROTEIN ASSAY PROTOCOLS
Protein assays are laboratory techniques used to measure the presence, concentration, or activity of proteins in a sample. These assays are crucial in various fields, including biomedical research, drug discovery, and diagnostics. Developing assay protocols is an essential step in…
STANDARD PROTOCOLS FOR LYSIS OF ESCHERICHIA COLI CELLS AND PARTITIONING OF CELLULAR COMPONENTS
Lysis of E. coli Cells: Harvesting the Cells: After the small-scale culture of E. coli reaches the desired optical density, harvest the cells by centrifugation. Resuspending the Pellet: Resuspend the cell pellet in an appropriate lysis buffer containing protease inhibitors…
PRODUCTION OF RNA AND PROTEIN SYNTHESIS
Production of RNA: RNA is produced through a process called transcription. The first step in this process is the initiation, where RNA polymerase binds to a specific region of DNA called the promoter. Then, the enzyme unwinds the DNA double…
DESIGN AND ASSEMBLY OF RECOMBINANT PLASMID CONSTRUCTS FOR PROTEIN EXPRESSION
In order to design and assemble ad hoc combinations of DNA precursors in silico and then in the laboratory to build recombinant plasmid constructs capable of directing expression of a recombinant protein, several steps and considerations need to be taken…
CELL COMPARTMENTALISATION AND PROTEIN TRANSPORT IN CELLS
Cell compartmentalisation refers to the organization of eukaryotic cells into distinct membrane-bound compartments called organelles. These organelles serve specific functions within the cell and are essential for maintaining cellular homeostasis. Proteins are transported between organelles through a complex network of…
EXPERIMENTAL TECHNIQUES FOR PROTEIN PURIFICATION AND CHARACTERIZATION
Protein purification and characterization are crucial steps in understanding the structure, function, and stability of proteins. There are several experimental techniques available for purifying and characterizing proteins, each with its own strengths and limitations. The choice of technique depends on…
PROTEIN FINGERPRINTING WITH SDS-PAGE
SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel electrophoresis) is a widely used technique for separating proteins based on their molecular weights. It is a crucial tool in protein fingerprinting, which involves the analysis of protein patterns to identify and characterize them. Principle…
PROTEIN FOLDS AND RESTRICTIONS IMPOSED BY PEPTIDE BOND PLANARITY
Proteins are composed of long chains of amino acids, and the way these chains fold into specific three-dimensional structures is crucial to their function. Protein folds are determined by the arrangement of secondary structural elements, primarily alpha helices and beta…
PRODUCTION OF PROTEIN LYSATES AND ANALYSIS OF PROTEINS BY WESTERN BLOTTING
Protein lysates are commonly used in molecular biology and biochemistry research to study the expression and function of proteins within cells. The process of producing protein lysates involves the extraction of proteins from cells or tissues, followed by their analysis…
HYPOTHESES REGARDING PROTEIN GLYCOSYLATION USING ARRAYS OR ONLINE BIOINFORMATICS TOOLS
To develop hypotheses regarding protein glycosylation using arrays or online bioinformatics tools, one can follow these steps: Data Collection: Gather relevant data on the protein of interest, including its sequence, structure, and any known glycosylation sites or patterns. Array Design:…
MECHANISMS OF ACTION OF PROTEIN SYNTHESIS INHIBITORS UNVEILED
Introduction Protein synthesis inhibitors are antibiotics that prevent bacteria from producing essential proteins needed for growth and survival. These antibiotics work by binding to the 30S ribosomal subunit, which is responsible for protein synthesis. The most common mechanism of action…
THE ESSENTIAL GUIDE TO GLOBULAR PROTEIN
Globular proteins are a type of protein with a characteristic three-dimensional structure that is compact and roughly spherical in shape. They play crucial roles in various biological processes, including enzymatic reactions, signaling, transport, and immune responses. The structure of globular…
PROTEIN PROCESSING AND TARGETING
Posttranslational modifications of proteins Posttranslational modifications (PTMs) are covalent modifications that occur on proteins after translation, altering their structure, stability, localization, and function. Here are some of the major types of PTMs: Phosphorylation: Addition of a phosphate group (PO₄³⁻) to…