A total of 29 WT chromodomains and two CBX1 mutants were expressed as glutathione S-transferase (GST) fusion proteins using pGEX-KG vector in Escherichia coli strain BL21(DE3) based on a previously described protocol (19). A total of 467 unmodified and modified peptides [selected by a bioinformatics pipeline previously developed and described in (19, 20) and in the Supplementary Methods and Materials] were synthesized by Sigma-Aldrich (desalted, mass spectrometry checked). The peptides were then printed as triplets onto glass slides (ArrayIt). The CBX1 mutant was screened against an Active Motif MODified Histone Peptide Array. Binding of the peptide microarray and the chromodomain proteins was performed and analyzed using a previously described protocol [see (19) and Supplementary Methods and Materials for details].

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.