SpCase9-MBP fusion protein purification

The homologous Cas9 was fused downstream of the Maltose binding protein and a TEV protease cleave site. For bacterial expression of the recombinant proteins Cas9, corresponding plasmid was transformed into BL21-Plys (DE3)- competent cells (Stratagene). Expression of the recombinant proteins was induced by subculturing 5 ml starter culture into 1000 ml LB, adding at OD600Continue reading “SpCase9-MBP fusion protein purification”

Converting cryosparc to relion

1 – run cryosparc2star in the directory of interest, with the .cs particle file. Make sure to use the —relion2 flag.  csparc2star.py DrmABDNAcryosparc_P251_J73_008_particles.cs cryosparc-particles.star –relion2 2 – change .mrc to .mrcs within star file  sed -i ‘s/.mrc/.mrcs/g’ p205_j279_particles.star 3 – Create new extract job directory corresponding to the one in the cryosparc-particles.star file (use viContinue reading “Converting cryosparc to relion”