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. 2010 Jun 21;2010:172484. doi: 10.1155/2010/172484

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Copyright © 2010 Naganath Mandi 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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(a) Restriction analysis of the PCR product of pPIC9K-rPA mutated constructs. M: lambda EcoR1/HindIII digest, lane 1: PCR product from N12P clone+ Sma1; lane 2: N278S clone+ XbaI; lane 3: N12P, N278S +Xba1/Sma1. (b) Integration PCR of Pichia recombinants carrying native rPA gene. M: lambda EcoR1/HindIII digest; lanes 1–4 geneticin resistant clones of Pichia carrying ppIC9K-reteplase gene and its mutants integrated into its genome. Lane 1: N12P clone; lane 2: N278S; lane 3; N12P, N278S; lane 4: WT reteplase.