Figure 8.

Retroviral expression of CMP transgenes with point mutations. (A) Constructs of CMP transgenes with point mutations. (1) Single mutations: CMP-D1, CMP-D2, and CMP-E1; (2) double mutations: CMP-D1D2, CMP-D1E1, and CMP-D2E1; (3) triple mutation: CMP-D1D2E1. X indicates the position where the mutation is made. (B) Expression of CMP transgenes by CEFs. Western blot of cell lysates from cultures of transfected CEFs. Seven and a half microliters of cell lysates were loaded under reducing conditions in each lane. The antibody used to detect CMP was 9E10, which detects the myc-tagged recombinant protein. Note the amount of synthesized recombinant CMP in cell lysates is similar in all of the constructs, including wild-type and mutants. Same results were obtained from conditioned medium from cultures of transfected CEFs.