Eukaryotic expression: developments for structural proteomics

Aricescu, A.R.; Assenberg, R.; Bill, R.M.; Busso, D.; Chang, V.T.; Davis, S.J.; Dubrovsky, A.; Gustafsson, L.; Hedfalk, K.; Heinemann, U.; Jones, I.M.; Ksiazek, D.; Lang, C.; Maskos, K.; Messerschmidt, A.; Macieira, S.; Peleg, Y.; Perrakis, A.; Poterszman, A.; Schneider, G.; Sixma, T.K.; Sussman, J.L.; Sutton, G.; Tarboureich, N.; Zeev-Ben-Mordehai, T.; Jones, E.Y.

doi:10.1107/S0907444906029805

Acta Crystallographica Section D
Acta Crystallographica
Section D BIOLOGICAL CRYSTALLOGRAPHY

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Figure 3
Co-expression of the His-Ama and Nrt proteins in P. pastoris. Cells harbouring both genes were induced for 2 d in BMMY medium. Proteins were analyzed on 12% SDS–PAGE followed by staining with GelCode (Pierce). Arrows indicate the predicted positions of the proteins. Lane 1, analysis of a 15 µl culture supernatant following 2 d induction; lane 2, proteins obtained upon elution from Ni–NTA agarose beads. Mass-spectrometric analysis revealed that the band at ∼45 kDa (lane 2) contains peptides from both Ama and Nrt.

BIOLOGICAL
CRYSTALLOGRAPHY

ISSN: 1399-0047

Volume 62| Part 10| September 2006| Pages 1114-1124

doi:10.1107/S0907444906029805

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