The ability to overexpress genes in microorganisms has revolutionized the field of synthetic biology. Gene overexpression involves the production of large quantities of target proteins by increasing the transcription and translation of the gene of interest. This technique has been applied in the engineering of microorganisms to produce a wide range of products, including biofuels, chemicals, and biopharmaceuticals.
There are several strategies for gene overexpression in strain engineering. One common method is gene synthesis, which involves the design and synthesis of genes optimized for expression in specific host organisms. Gene experts modify the codon usage, remove destabilizing sequences, and add regulatory elements to enhance expression. Therefore, the target expression level of the gene is achieved through the modification and design of the gene.
Another approach is cloning. Experts utilize state-of-the-art techniques, such as restriction enzyme digestion and DNA ligation, polymerase chain reaction (PCR) amplification, plasmid-mediated transformation, and gene transfection, to clone synthesized genes into different expression vectors, enabling high levels of gene expression.
In some cases, to enhance gene expression, regulatory elements that may reduce gene expression levels can be identified and removed. This type of gene optimization can be achieved by modifying the promoter, ribosome binding site, and other regulatory elements to obtain optimal gene expression levels.
Overall, gene overexpression is a powerful tool in strain engineering to enhance the production of specific compounds or metabolic activities. It involves increasing the expression level of specific genes through techniques such as gene synthesis, cloning, gene optimization, and protein expression. From a synthetic biology perspective, researchers can explore the role of gene overexpression in strain engineering and how they can optimize gene overexpression for diverse applications.