Abstract
The application of the peptide-linked β2-microglobulin (β2m) strategy is limited in some cases due to the incompatibility between the sequences of the peptides and the restriction sites of the plasmid vectors. An isocaudamer technique was adapted to overcome this restriction. Three peptide-linked β2m genes, HBc18–27-hβ2m gene, OVA257–264-mβ2m gene and HER2/neu369–377-mβ2m gene, were inserted into the pET28a vectors with this technique. The corresponding proteins were expressed in Escherichia coli with yields of over 50 mg/l culture and purities of over 80%. This strategy facilitates the construction of peptide-linked β2m molecules and will simplify the preparation of major histocompatibility complex-peptide complexes.
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This work was supported in part by the grants from the National Nature Science Foundation of China (30571703) and National Science Fund for Distinguished Young Scholars of China (30325017).
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Meng, F., Shen, C., He, Y. et al. Cloning and expression of three peptide-linked β2-microglobulin molecules in Escherichia coli with an isocaudamer technique. Biotechnol Lett 31, 831–836 (2009). https://doi.org/10.1007/s10529-009-9950-3
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DOI: https://doi.org/10.1007/s10529-009-9950-3