E.coli is the preferential one of bacterial systems for recombinant protein production and large scale fermentation as its advantages including fast rate of reproduction, ease of culture, and rich knowledge about its genetics. Bacterial?protein expression service offered by DetaiBio provide a relatively simple, fast, and inexpensive platform for native and heterologous recombinant protein production. Contact us for quotations and more information.
|Plasmids or sequences of target protein||
For more detailed information, please contact us.
Rare codon may cause the failure of protein expression and affect the secondary structure of mRNA. After performing thousands of projects, we can analyze and optimize the codon of the sequence of target protein perfectly by using our unique bio-informatics software MaxcodonTM.
We can start the gene synthesis after codon optimization. Meanwhile, dozens selections of expression vectors can be selected, including pET series (pET28a, pET30a, pET43.1 a, etc.), pGEX4T-1, pBAD24, pGADT7, pQE-30, etc. If you do not know how to select, we are happy to assist you.
According to the property of target protein, DetaiBio can also help to choose the most suitable competent cells, which include several kinds, such as BL21 (DE3), OrigamiB (DE3), Rosetta (DE3), BL21 CodonPlus (DE3), ESL, etc. Then we perform pilot expression tests and explore the best expression conditions.
According to expression level and customer’s specific needs, we scale up the protein expression with suitable level using the optimized conditions. We can purify the target protein with major protein purification methods (such as affinity chromatography, size-exclusion chromatography, and hydrophobic interaction chromatography) using advanced AKTA purification equipment.
Target protein will be detected by SDS-PAGE and western blot. Then target protein will be delivered with the QC report.
If your proteins did not express in e.coli system, the most possible reason is that its gene is from eukaryotes. Eukaryotic genes generally have signal or leader sequences and intervening sequences, which can influence the result of protein expression in bacteral system. In addition, E.coli cells lack the post-translational modification system of eukaryotic proteins, which may result in failure of eukaryotic proteins expression that require glycosylated phosphorylation.
You can replace the host bacteria,such as BL21 Condon plus (DE3) or Rosetta, which is specially designed to express eukaryotic proteins.
Several points need your attention, including: translation initiation site, GC content, secondary structure of mRNA, codon, plasmid vector, the size of target protein and gene mutation.
You can analyze the gene sequence of target protein and optimize the culture conditions (such as medium composition, temperature, induction time, concentration of inducer, and induction time).
For quote, please fill in the online form and email to Sales@Detaibio.com.
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