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. 2010 Jun 27;2010:376927. doi: 10.1155/2010/376927

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Copyright © 2010 Muriel Roulet et al.

This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Expression of α1(V) without the native signal sequence and the C-propeptide (a, b) and of α1(V)/α2(V) chains (c) in P. pastoris. (a) 5% SDS-PAGE analysis under reducing conditions of the soluble fraction of acidic extraction from untransformed yeast cells (lane 1) and from yeast cells transformed with the construct pPICZα-C5A1N (lane 2). (b) Western blot analysis of the same samples as in panel (a) probed with monoclonal pan-collagen antibody 95D1A. (c) Western blot analysis of the soluble fraction of neutral extraction of yeast cells transformed with α1(V) and α2(V) chains and induced with 0.5% methanol induction. Samples were incubated without (lane 1) or with (lane 2) pepsin before analysis. Left, molecular mass standards expressed in kDa.