Biochem Exam 5

Restriction Endonucleases

Bacterial enzymes that cleave DNA at specific sites, recognizing palindromic sequences and creating fragments.

Nucleic Acid Fragment Separation

Separation of DNA fragments using agarose gels and visualization with dyes like Ethidium bromide.

DNA Cloning

Process of making identical copies of a piece of DNA by inserting it into a vector plasmid and introducing it to bacteria.

Plasmids

Circular DNA separate from the genome, capable of autonomous replication and containing antibiotic resistance genes.

Eukaryotic gene expression in bacteria

Eukaryotic genes need to be converted to complementary DNA (cDNA) before expression in bacteria due to differences in promoters and the presence of introns.

Amplifying DNA by PCR

Polymerase Chain Reaction (PCR) uses DNA primers, DNA polymerase, and thermal denaturation to amplify target DNA.

Proteome

The set of proteins expressed and modified in a cell, which varies from the genome.

Subcellular Fractionation of tissue

Process of homogenization and differential centrifugation to separate cellular components.

Protein Purification

Separation of proteins based on solubility, charge, size, and specific binding activity using methods like chromatography.

Salting out

Precipitation of proteins at high salt concentrations, followed by centrifugation and resuspension.

Dialysis

Removal of small molecules from a protein solution by diffusion through a semipermeable membrane.

Column Chromatography

Separation of protein mixtures over a solid phase using a liquid phase to mobilize the proteins.

Ion Exchange Chromatography

Separation of proteins based on charge using cation or anion exchangers.

Affinity Chromatography

Separation of proteins based on binding affinity using a ligand solution.

Size Exclusion Chromatography

Separation of proteins based on size using a column containing cross-linked polymers.

Electrophoresis for Protein Analysis

Separation of proteins based on charge and size using an electric field and gel matrix.

Signal Transduction Pathways

Process by which extracellular signals are amplified and converted to cellular responses.

Basic Types of Receptors

G protein-coupled receptors (GPCRs), receptor tyrosine kinases, and gated ion channels.

G Protein-Coupled Receptors (GPCRs)

Integral membrane proteins that bind signal molecules and activate G proteins to initiate intracellular signaling.

Receptor Tyrosine Kinases

Membrane-bound proteins with tyrosine kinase activity that phosphorylate target proteins in response to ligand binding.

Gated Ion Channels

Regulate the transport of ions across the cell membrane in response to changes in membrane potential or ligand binding.

Insulin Pathway

Insulin binds to its receptor, leading to a cascade of protein-protein interactions and enzymatic reactions that regulate glucose transport.

Steroid Hormones

Oxidized derivatives of sterols synthesized from cholesterol, carried in the bloodstream, and involved in male and female sex hormone regulation.