Recombinant DNA Technology in Vaccine and Insulin Production
Recombinant DNA Technology in Vaccine Manufacture
Steps for Recombinant DNA Preparation
1. Isolation of Genetic Material or DNA
Genetic material is present inside the cells. It has to be obtained in pure form without even the attached histones and other proteins. To get the desired DNA, cells are treated with lysozyme (for bacteria), cellulase and pectinase (for plant cells), and chitinase (for fungal cells).
- The treated cells are homogenized and centrifuged to rupture the cells as well as nuclear envelopes.
DNA Restriction Digestion and Agarose Gel Electrophoresis
Understanding Restriction Enzymes
Restriction enzymes (RE), or restriction endonucleases, act as molecular scissors by cutting DNA at specific recognition sequences. These enzymes are isolated from various cellular strains and typically recognize palindromic sequences of 4 to 6 base pairs. Once the target site is located, the enzyme cleaves both strands of the DNA double helix, resulting in restriction fragments.
Common Restriction Enzymes
Every enzyme has unique target sites. For example, Lambda DNA
Read MoreDrug Physicochemical Properties and Biological Action
Physicochemical Properties in Relation to Biological Action
Physicochemical Properties refer to the physical and chemical characteristics of a drug molecule that influence its behavior in biological systems. These properties play a crucial role in determining the drug’s absorption, distribution, metabolism, and excretion (ADME). They include:
- Ionization
- Solubility
- Partition Coefficient
- Hydrogen Bonding
- Protein Binding
- Chelation
- Bioisosterism
- Optical and Geometrical Isomerism
1. Ionization
Ionization is a key
Read MorePharmacology Drug Classes and Mechanisms Summary
NSAIDs: Pain Relief and Inflammation Reduction
NSAIDs (Non-Steroidal Anti-Inflammatory Drugs) are a class of meds that reduce inflammation, pain, and fever. They’re like the ultimate pain relief squad.
Classification
- Non-selective COX inhibitors: Aspirin, Ibuprofen, Diclofenac, Naproxen
- COX-2 selective inhibitors: Celecoxib, Etoricoxib
Pharmacology of Aspirin
- Mechanism: Aspirin irreversibly inhibits COX-1 and COX-2 enzymes, reducing prostaglandin synthesis, which leads to decreased pain, inflammation,
Molecular Muscle Physiology and Anti-Doping Regulations
Cellular Fate and the Cell Cycle
Cells can follow four major fates: proliferation (cell division), quiescence (reversible resting state), differentiation (specialization with reduced division), or apoptosis (programmed cell death). These outcomes depend on:
- Internal factors: Gene expression patterns, regulatory proteins (e.g., p53, pRb), and metabolic status.
- External factors: Growth factors, hormones, nutrient availability, and cell–cell contact.
Cell Cycle Mechanics
- G1 phase: Cell growth, normal
Animal
Applications of cell culture
Cell culture is one of the major tools used in cellular and molecular biology, providing excellent
model systems for studying the normal physiology and biochemistry of cells (e.G., metabolic
studies, aging), the effects of drugs and toxic compounds on the cells and mutagenesis and
carcinogenesis. It is also used in drug screening and development and large scale manufacturing
of biological compounds (e.G., vaccines, therapeutic proteins). The major advantage of using cell
culture
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