E. coli is one of the most well-known and utilized host cells in protein production systems and services. Researchers and scientists all over the globe rely on this molecular technique for its simplicity, affordability, and high-concentration cultivation yields. We’ll discuss a few critical factors affecting E. coli expression and offer optimization insights.
Purification methods for E. coli expression can vary, but they are significant elements for a complete service. The chosen method will depend on the protein in service and the type of host cell, whether eukaryotic or prokaryotic.
Because E. coli generates higher yields, researchers prefer this expression system of interest over others when applicable. They can simplify purification measures by attaching a fusion tag to the designated protein. The tag’s composition can influence the solubility and expression of the protein in interest. Common tags include the following:
Common post-translational modifications (PTM) include N-Acetylation, S-Nitrosylation, Ubiquitination, Glycosylation, and Phosphorylation. When compared to eukaryotic proteins, there are a fewer number of post-translational modifications with bacterial proteins.
Plasmids are small, stable microbes that are easily manipulated. When a lab designs a plasmid, they are known as vectors, allowing labs to insert genes of interest and conduct an expression service. Common vectors used for E. coli expression include pSumo, pET, pGEX, and pBAD. Plasmids play a fundamental role in genetic transfer processes, leading to a better adaptation regardless of environmental diversities for the host bacteria.
The primary objective of codon optimization is to improve the expression of the proteins without altering the amino acid's sequencing. Typically, codon optimization reduces the complexity of a specific recombinant gene.
During the protein synthesis phase, the genetic codes translate genetic data from mRNA and DNA into proteins. The nucleotide triplets, or codons, declare which amino acids are necessary for the protein synthesis phase while upholding the existing amino acid sequence.
The host strain is one of the most critical factors affecting E. coli expression. It plays a role in overall expression conditions and times. Each host strain carries a genotype and a set of key features, but the general use or each host will determine the best option. The larger the collection of hosts, the more optimized an expression service. The lab can screen to find the most suitable solution for a project. A few common host strains of E. coli include the following:
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