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. 2004 Aug;16(8):1979–2000. doi: 10.1105/tpc.104.023614

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Copyright © 2004, American Society of Plant Biologists

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Figure 8. — Purification, Cloning, and Expression of PSRP1.

(A) Purification of the 27-kD PSRP1 from pumpkin phloem sap using a combination of Q-Sepharose and metal chelation chromatography. Protein profiles contained within the anion-exchange fractions (L, loading; FT, flow-through; E, elution) were resolved by 10% SDS-PAGE (an equal volume [20 μL] was loaded for each fraction). Protein profiles for metal chelation resolved as above (FT, flow-through; W1/ 2, washes).

(B) Conceptual translation of CmPSRP1 (GenBank accession number AY326308) yielded a 20,454-D protein. The predicted molecular mass was consistent with the value of 21,004 D as determined by mass spectroscopy with the 27-kD PSRP1 in (A).

(C) Purification of recombinant (R)-PSRP1 using metal chelation chromatography. Proteins from each step were resolved by 10% SDS-PAGE and visualized using GelCode Blue.