Restriction Enzymes: Function, Types, and Biotechnology Applications
What Are Restriction Enzymes?
A restriction enzyme is a protein produced by bacteria that cleaves DNA at specific sites. This specific location is known as the restriction site.
Restriction enzymes are important tools for genetic engineering. They can be isolated from bacteria and used extensively in laboratories.
Bacterial Defense Mechanism
Restriction enzymes protect living bacteria from bacteriophages (viruses that infect bacteria). They recognize and cleave the bacteriophage’s DNA at its restriction
Read MoreEssential Spectroscopic Methods: NMR, IR, UV/Vis, and MS Applications
Fundamentals of Spectroscopy
Spectroscopy is the study of the interaction of electromagnetic radiation with matter, providing valuable information about molecular structure, composition, and properties. Different types of spectroscopy target various regions of the electromagnetic spectrum and yield specific data.
Major Spectroscopic Techniques
A. Nuclear Magnetic Resonance (NMR) Spectroscopy
- Principle: NMR is based on the absorption of radiofrequency radiation by nuclei with a non-zero spin (e.g., 1H,
E. coli Expression Systems: Host Selection, Vectors, and Industrial Titers
Escherichia coli Expression Systems
Advantages of Microbial Expression Systems
Bacterial and yeast systems share several advantages over mammalian expression systems:
- Higher growth rate and cell densities.
- Robustness at the cellular level.
- High expression levels.
- Simple cultivation and are easy to scale-up.
The selection of the appropriate expression system is determined by the characteristics of the target protein or product.
Why E. coli Dominates Microbial Expression
E. coli has dominated microbial expression
Read MoreCellular Transport Mechanisms and Concentration Gradients
Solution Concentration and Particle Movement
Many essential chemical reactions take place in the cytoplasm and aqueous solutions.
- Solution: A solute dissolved in a solvent.
- Concentration: The amount of solute in proportion to the amount of solvent in a solution.
- All particles in a solution possess kinetic energy, resulting in continuous random movement.
The Principle of Diffusion
Molecules and ions in a fluid are in continuous, random motion due to their kinetic energy. Diffusion is the net movement of
Read MorePlant Tissue Culture Technology: Methods and Applications
Introduction to Plant Tissue Culture (PTC)
Plant tissue culture (PTC), developed around the 1950s, was a significant addition to plant breeding methods. Conventional breeding techniques often struggled to meet the required demand for crops, making tissue culture a major advancement in breeding practices.
PTC is defined as the in vitro aseptic culture of cells, tissues, or whole plants under controlled nutritional and environmental conditions, primarily used to produce clones of plants.
The Principle
Read MoreBioprocess Scale-Up & Scale-Down: Industrial Optimization
Bioprocess Scaling Up: Lab to Industrial Production
Bioprocess scaling up is the methodology used to develop an industrial bioprocess from knowledge acquired at the laboratory scale, often based on mathematical and physical models.
Approaches to Bioprocess Scaling Up
Scaling up can be approached in two primary ways:
- Increasing the number of reactors: This method carries no risk of scaling errors since all reactors are identical. However, it incurs significant expenses for monitoring and managing numerous