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Today, we're discussing what happens once we’ve successfully inserted foreign DNA into a host cell. Can anyone tell me why expressing this DNA is essential?
We need to express the DNA to produce proteins that can be useful, right?
Exactly! Our goal is often to produce a specific protein encoded by that inserted DNA, known as a recombinant protein. Let's remember this by thinking of it as 'Expressing to Impress'.
What kind of proteins are we considering here?
Great question! Recombinant proteins can include enzymes, hormones, or even antibodies used in therapies. Now, after insertion, we need to optimize conditions for gene expression. Can anyone suggest what factors we need to consider?
Factors like temperature and pH?
Yes, temperature, pH, and the right nutrients are crucial for optimal protein yield. Now let's summarize: after inserting foreign DNA, we aim to express that gene under specific conditions to produce desirable proteins.
Having discussed the principles of gene expression, let's move on to large-scale production. Why do you think we can't just keep making proteins in tiny lab cultures?
Because we would need a lot more of the protein for real-world applications?
Correct! To meet demands, we require bioreactors—large vessels designed for this process. They provide the necessary conditions to cultivate organisms efficiently. Can anyone list some advantages of bioreactors?
They offer better temperature control and oxygen supply!
Well done! Bioreactors can also be adjusted for stirring, mixing, and nutrient input continuously, which is crucial for maintaining cell growth. Remember, our approach can influence yields significantly.
Is it true that the size of the bioreactor affects the yield?
Absolutely! Larger bioreactors can maintain high cell densities and, consequently, higher product yields. To sum it up, bioreactors are key for large-scale recombinant protein production.
After producing the desired proteins, what do you think should be the next step?
We need to purify and process them before they can be used.
Exactly right! This is referred to as downstream processing, which includes purification steps, formulation, and quality control. Can anyone outline why each of these is important?
Purification ensures that we have the actual product without contaminants, right?
Correct! If we don't purify, the product may not work as intended or could even cause harm. Quality control is just as crucial to ensure every batch meets the strict requirements—especially for medical applications. Remember, a product that doesn’t meet standards can be dangerous!
So, both the purification and quality control are vital in the entire biotechnology process?
Absolutely! The success of biotechnological applications relies heavily on effective downstream processing and maintaining high standards. Let's wrap up! We've learned that producing recombinant proteins involves effective gene expression, large-scale cultivation in bioreactors, and thorough downstream processing.
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The section elaborates on how, after inserting alien DNA into host cells, the next goal is to express that DNA to produce desired proteins, focusing on techniques for large-scale cultivation of these proteins using bioreactors.
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When you insert a piece of alien DNA into a cloning vector and transfer it into a bacterial, plant or animal cell, the alien DNA gets multiplied. In almost all recombinant technologies, the ultimate aim is to produce a desirable protein. Hence, there is a need for the recombinant DNA to be expressed.
This chunk introduces the concept that the primary goal of recombinant DNA technology is to produce proteins. When foreign DNA is inserted into a host organism (like bacteria or plants), it not only gets replicated but is also expected to be expressed to yield functional proteins necessary for various applications. Understanding how to induce this gene expression is critical for biotechnological applications.
Imagine a bakery where foreign ingredients (like chocolate chips or nuts) are added to a basic cookie dough recipe. The goal is to produce delicious cookies (the desirable protein) from that dough (the cloning vector) once these foreign ingredients are mixed in. Just like bakers must find the right temperature and baking time to make perfect cookies, biotechnologists must optimize conditions to express the foreign gene effectively.
After having cloned the gene of interest and having optimised the conditions to induce the expression of the target protein, one has to consider producing it on a large scale.
Once the gene of interest is successfully cloned into the host organism and the expression conditions are fine-tuned, the focus shifts to large-scale production of the protein. This means that scientists must carefully control the environment in which the host cells grow, ensuring they produce as much of the target protein as possible.
Consider a company that starts small but wants to grow its production line. Initially, they optimize the recipe for their popular snack, but then they need to scale it to produce tens of thousands of bags every day. They’ll have to control everything from ingredient quality to machine efficiency, akin to a lab controlling temperature and nutrient supply to maximize protein yield.
The cells harbouring cloned genes of interest may be grown on a small scale in the laboratory. The cultures may be used for extracting the desired protein and then purifying it by using different separation techniques.
This chunk explains that initial cultures of the genetically modified cells can be small, enough for laboratory-scale experiments. These cultures provide a way to test and extract the desired protein, which may undergo various purification processes to isolate it from other cellular components.
Think of brewing tea. You start with a small pot, allowing you to test the flavor. Once you perfect it, you can brew a larger batch. Similarly, in recombinant biotechnology, small-scale cell cultures allow scientists to test and refine the protein extraction process before moving to larger-scale production.
The cells can also be multiplied in a continuous culture system wherein the used medium is drained out from one side while fresh medium is added from the other to maintain the cells in their physiologically most active log/exponential phase.
This chunk discusses the significance of bioreactors in maintaining optimal conditions for cell growth. A continuous culture system allows nutrients to be refreshed while waste products are removed, leading to higher yields of the target protein as cells remain in their most productive phase.
Consider a hydroponic garden where plants have a steady supply of nutrients and oxygen. This ensures the plants grow optimally, just like cells in a bioreactor thrive with continuous medium renewal, maximizing protein production for industrial use.
Thus, bioreactors can be thought of as vessels in which raw materials are biologically converted into specific products, individual enzymes, etc., using microbial plant, animal or human cells.
Bioreactors are designed to provide ideal conditions for the growth of cells used in biotechnology, facilitating the transformation of raw materials into desired products, including proteins. They are equipped with systems to monitor and control conditions such as temperature, pH, and oxygen levels.
Imagine a controlled environment like an aquarium where conditions such as water temperature and filtration are carefully managed to foster fish growth. Similarly, a bioreactor maintains specific conditions to maximize the growth and productivity of the cells within it.
A stirred-tank reactor is usually cylindrical or with a curved base to facilitate the mixing of the reactor contents. The stirrer facilitates even mixing and oxygen availability throughout the bioreactor.
This concluding chunk wraps up the section by describing a common type of bioreactor, the stirred-tank reactor. Its design ensures optimal mixing and oxygen transfer, which are crucial for the effective growth of cells and production of proteins.
Think of making a smoothie. You blend fruits, yogurt, and ice to ensure everything combines evenly; a stirred-tank reactor does the same for cells and nutrients, ensuring they mix thoroughly for efficient growth and protein production.
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Key Concepts
Gene Expression: The process of translating an inserted gene into a protein.
Bioreactors: Large-scale vessels used for cultivating organisms to produce proteins.
Downstream Processing: The purification and preparation of the recombinant product for end use.
Selectable Marker: Gene used for selecting transformed cells.
See how the concepts apply in real-world scenarios to understand their practical implications.
Insulin produced through recombinant DNA technology is used in diabetes management.
Human growth hormone produced in bacteria for therapeutic use.
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In the bioreactor, we mix and stir, to grow our cells, in a controlled blur.
Imagine a farmer (the researcher) cultivating a field (the bioreactor) to grow crops (proteins) that will feed the community (therapeutic applications). Each season (batch process), he must ensure he waters them correctly (maintains conditions).
PEAR: Protein Expression in Amplified Reactors - to remember the steps of protein production.
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Term
Recombinant Protein
Definition
Bioreactor
Downstream Processing
Selectable Marker
Review the Definitions for terms.
Term: Recombinant Protein
Definition:
A protein produced from the expression of a cloned gene in a host organism.
Term: Bioreactor
A vessel that provides a controlled environment for the cultivation of cells or microorganisms.
Term: Downstream Processing
The series of processes that purify and prepare the bioproduct for end use after it has been synthesized.
Term: Selectable Marker
A gene used to identify cells that have successfully taken up foreign DNA.
Flash Cards
Glossary of Terms