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Did you know? This is how the world of genetic transcription works!
When we explore the mysteries of life, genes represent the blueprint of life, and the process of gene transcription is the key to how this blueprint is read and realized. Recent research shows that heterologous expression technology opens a window into the inner workings of this cell, allowing scientists to express specific genes in non-natural host tissues. This process is achieved through recombinant DNA technology. .
Heterologous expression not only provides a simple and effective means to study molecular function, but also allows us to detect interactions between mutations and proteins, which may not be observed in nature.
Heterologous expression is common in a variety of host systems such as bacteria, yeast, mammalian cells, and plant cells. The choice between these host systems depends on a variety of factors, including production cost, expression level, and required post-translational modifications. The inner workings of cells are complex processes, and heterologous expression allows us to study how various gene combinations affect protein function and interactions.
Technical Overview
Heterologous expression technology can be divided into two main parts: gene isolation and host entry technology.
Technology for isolating specific genes
Gene identification can be carried out through computer-assisted methods, which are called heterologous screening techniques. For example, the cDNA sequence digital library integrates data from many gene sequencing projects and provides convenient access to known gene sequences. If the genomic sequence of the target gene is unknown, experimental methods such as random fragmentation, cloning, and screening can be used to determine its phenotype. In this process, the identification of restriction enzymes is particularly important, because these enzymes can cut DNA according to specific sequences and help scientists extract the desired genes.
Technology for integrating genes into hosts
When introducing genes into host cells, researchers can choose from a variety of methods, such as gene gun delivery, electroporation, viral transduction and lipofection. Each of these methods has advantages and disadvantages and can meet different experimental needs.
For example, the physical properties of gene guns can reduce immune responses and are suitable for gene delivery in plant and animal cells. Electroporation uses high voltage to create instantaneous holes, allowing DNA to enter cells, and can be applied to almost all tissue types.
Host system selection
Selecting an appropriate host system is one of the keys to successful gene heterologous expression research. Common host systems include Escherichia coli (E. coli), yeast (such as S. cerevisiae and P. pastoris), insect cells, etc.
E. coli
E. coli is a widely used host due to its rapid growth (every 20-30 minutes), low cost and rich genetic knowledge. However, it should be noted that when expressing certain proteins, it may cause precipitation and aggregation, which requires additional refolding and recovery processing.
Yeast
Yeast can assist in post-translational modification of proteins, especially for the production of human drugs. These microorganisms can be cultured relatively cheaply and grow rapidly. However, production costs and nutrient requirements are relatively high, making it a technical challenge to strike a balance between different host systems.
Mammalian cells
Although the culture of mammalian cells is more difficult and expensive than other systems, it has more advantages in post-translational modification. For proteins that require clinical application, mammalian cell lines are the most ideal host system.
Application
Heterologous expression technology has a wide range of applications and can be used in biomolecule research, drug development and other fields. For example, the heterologous expression of nitrogen enzymes helps to understand the physiological processes inside cells and supports drug development from the genetic level, which will occupy an important position in future medical technology.
For example, the first heterologous protein product launched on the market was human insulin (Humulin), which was produced using a certain strain of E. coli and successfully shaped the early development of genetic engineering technology.
With the deepening of research and the advancement of technology, heterologous expression will surely promote the development of biotechnology and provide unlimited possibilities for future disease treatment, drug development and genetic research. Have you also thought more deeply about the role that gene transcription plays in life?
