Figure 2
Construction of the MK5L 337Aexpression plasmid for microinjection. (A) Expression vector for the delivery of the transgene. The injected fragment from the vector contains the CMV promoter, the SV40 intron, cDNA for HA-tagged MK5L 337Aand an SV40 polyA tail (see Methods). The blue arrows indicate in which restriction site (Sph I) the vector was linearized. Panels B to D illustrate genotyping and expression of MK5L 337Awith : (B) β-globin presence as a control for the DNA; (C) transgene integration as shown by PCR on genomic DNA. In panels B and C: lane 1 is the positive control (Ctrl) consisting of DNA from a TG mouse; lane 2, lkb+ ladder (L); lane 3, non-transgenic offspring, (NTG); lane 4, transgenic offspring (TG). (D) Transgenic RNA expression: in lane 1, WT C57BL/6; lane 2, mouse line A; lane 3, mouse line B transgenic offspring and lane 4, mouse line B non-transgenic littermate. The left lane in panel D represents the DNA size marker.