Tissue-specific expression of mKng1 and mKng2. (A) RT-PCR of mKng1- and mKng2-derived HK mRNA transcripts in murine tissues. PCR products were resolved using 0.9% agarose gel electropheresis, and stained using ethidium bromide. Arrows indicate the location of PCR products from mKng1- and mKng2-derived HK mRNA. The lane on the left side of the gel (M) contains a DNA ladder. GAPDH mRNA was amplified in parallel to control for the DNA content of individual samples. L indicates liver; Lu, lung; K, kidney; A, adrenal; Bl, bladder; Br, brain; Mu, muscle; Thy, thymus; Sp, spleen; and H, heart. (B) In vitro translation of the regions of liver- and kidney-derived cDNA encoding Val₃₆₄ to Arg₅₄₀ of mKng1 (and corresponding region of mKng2). cDNA was amplified using the primers depicted in Figure 3B. PCR products were cloned into pBluescript II KS⁺, and used to synthesize capped mRNAs. In vitro translation was performed in the presence of ³⁵S-methionine, and labeled polypeptides detected using 15% SDS-PAGE and autoradiography. As demonstrated in this figure, liver cDNA yielded a slightly larger polypeptide than that derived from kidney, consistent with expression of mKng1-derived HK mRNA in the former, and mKng2-derived HK mRNA in the latter.