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The A175T variant lacks KDS reductase activity. (A) Expression of FVT1 and fvt1-A175T in E. coli; SDS/PAGE of uninduced (−) and induced (+) whole-cell lysates. (B) Purification of the His6 fusion protein. Both the Thr-175 variant from an affected calf and the normal Ala-175 are expressed and purified in parallel under the same conditions with comparable results. (C) In vitro KDS reductase assay of the protein fractions shown in B.
