Abstract
Human ribosomal proteins play important structural and functional roles in the ribosome and in protein synthesis. An efficient method to recombinantly produce and purify these proteins would enable their full characterisation. However, the production of human ribosomal proteins can be challenging. The only published method about the recombinant production of human ribosomal proteins involved the recovery of proteins from inclusion bodies, a process that is tedious and may lead to significant loss of yield. Herein, we explored the use of different Escherichia coli competent cells and fusion protein tags for the recombinant production of human ribosomal proteins. We found that, by using thioredoxin as a fusion protein, soluble ribosomal protein could be obtained directly from cell lysates, thus leading to an improved method to recombinantly produce these proteins.
Highlights
Several high-resolution structures of the ribosome have become available by using X-ray crystallography and cryo-electron microscopy[8,9,10,11,12]
As the gene sequences of human ribosomal proteins typically contain rare codons that could slow down or impede the regular mRNA translation process in Escherichia coli[18], the use of E. coli competent cells that contain a higher level of tRNAs that recognise rare codons were explored
In order to minimise the variables in this study, we chose four human ribosomal proteins that are of similar size, isoelectric point and amino acid composition; They are S10, S15, S18 and L11 (Tables 1 and 2)
Summary
Several high-resolution structures of the ribosome have become available by using X-ray crystallography and cryo-electron microscopy[8,9,10,11,12]. Recombinant production of human ribosomal proteins with poly-histidine tag. The resulting plasmids www.nature.com/scientificreports were introduced into E. coli BL21 (DE3) competent cells and protein expression trials were conducted to find the optimal growth condition to obtain soluble ribosomal proteins from the cell lysate.
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