THINGS TO CONSIDER FOR YOUR PROJECT
- Sequence Optimization: In highly expressed genes, a narrow set of codons is used and these codons correspond to the more abundant tRNA species. This minimizes the risk of tRNA depletion during translation. In fact, the codons in a gene may be true bottlenecks, especially in cases where foreign genes are expressed in a host in which the usage of codons in highly expressed genes does not resemble the usage of codons in the species from which the foreign gene originates. In such cases, it has been shown that substitution of rare codons in the introduced gene may increase the yield dramatically. In addition, replacement of rare codons might decrease the chance of misincorporation and protect the protein from premature turnover.
- Expression System: In order to decide which host is most suitable the amount and the degree of purity of the product as well as its biological integrity and potential toxicity should be considered. For example, bacterial expression systems are not suitable if post-translational modification is required to produce a fully functional recombinant product.
- Expression Vector: Once the host has been selected, many different vectors are available for consideration. However, as for the selection of a suitable host system, the final choice of vector should take into consideration the specific requirements of the application and will, of course, be influenced by the behaviour of the target protein.
- Tag: As for the selection of host and vectors, the decision to use which tag must be made according to the needs of the specific application. The two most commonly used tags are glutathione S-transferase (GST tag) and 6 x histidine residues ((His)6 tag).
Upon receiving your sequence information, the Bio Basic scientists will analyse the sequence to determine characteristics like molecular weight, pI, solubility, susceptibility to oxidation and hydrophobicity. Every protein is different, there is no standard way for protein expression and purification, and the protocols and strategies must be worked out for each individual protein and with its intended use in mind. Scientists at Bio Basic use their experience and expertise in purifying recombinant protein to best tailor the service for your research needs.
Contact your BaseClear account manager using our contact form to discuss your protein expression and purification project!