FIG. 9.
Identification of the chicken ggPRX gene promoter. (A) Alignment of the nucleotide sequences of the human -14 gene promoter region (numbers correspond to GenBank sequence AE006462) and of the CpG island from the upstream region of the chicken α-globin domain (numbers correspond to GenBank sequence AF098919). The regions sharing sequence identity are shaded. (B) Transient transfection experiments in HD3 cells (left) and HeLa cells (right); the constructs are shown on the far left and CAT activity is shown on the right. As a negative control, the promoterless pCAT3 basic vector (Promega) was used, and a construct with the erythroid cell-specific chicken αD-globin promoter driving CAT gene expression was used as a positive control in HD3 cells and as a negative control in HeLa cells. CAT activity was normalized, considering the efficiency of transfection. Data are the average of three independent experiments, and error bars show the SEM. (C) RT-PCR analysis of ggPRX gene transcription in HD3 cells. (Top) Positions of the regions examined on the map of the upstream part of the chicken α-globin gene domain. (Bottom) Amplification of test regions 1 to 4. Lanes marked “−RT” were loaded with products of direct PCR amplification of RNA (without RT polymerase); lanes marked “DNA” were loaded with products of amplification of 100 ng of chicken genomic DNA.