Research Paper Volume 3, Issue 3 pp 237—261

Figure 1. Diagram of transgenic constructs. (A) The “Tet-on” conditional transgene expression system. The rtTA transgenic construct (or “driver”) contains the tissue-general actin5C promoter driving expression of the artificial transcription factor rtTA. The target constructs were generated by cloning the indicated cDNA fragments downstream of the DOX-inducible promoter in the USC1.0 vector between the unique PstI and EcoRI sites. The number of bases present upstream and downstream of the A residue of the ATG start codon for normal translation are indicated for each cDNA insert. The rtTA protein will bind to the 7 Tet-O sites in the target construct promoter and activate transcription only in the presence of DOX. (B) Diagram of the hUbb construct. The number 1 indicates the A of the normal ATG start codon for translation of hUbb, and the stop codon is indicated by a black asterisk. (C) Diagram of the hUbb⁺¹ construct. The GAGAG hotspot for MM is indicated in blue, and the GT dinucleotide is indicated in purple. Note that in the hUbb⁺¹ construct the GT dinucleotide has been deleted so that this construct constitutively encodes hUbb⁺¹ protein. The amino acid sequence of the peptide used to generate the hUbb⁺¹ antibody is indicated in red. The independently derived transgenic strains are given names comprised of the name of the inserted construct (e.g., hUbb or hUbb⁺¹) followed by a unique number in brackets indicating the particular independent transgenic line.