Transcription start sites of human HSD17B10 gene were determined by primer extension. Part A: (M), DNA molecular standards of which the sizes (bp) are indicated; (1), controls run without reverse transcriptase; (2), without RNA; (3), without primer; and (4), extension reaction mixture analyzed in parallel on an 8% polyacrylamide gel. The dideoxy sequencing ladder is included in the Additional file 2. Part B: Two major transcription start sites at -37 and -6, respectively, and a minor one at -12 are indicated by vertical arrowheads a, b, and c, respectively. CpG dinucleotides in the proximal promoter and the HSD17B10 cDNA nucleotide sequence are in bold. The oligonucleotide primer used in this study was complementary to a fragment of the HSD17B10 cDNA nucleotide sequence, and is shown by a long dash arrow. Both the initiation codon ATG, numbered as +1, and the CCAAT box are underlined.
Yang et al. BMC Biochemistry 2013 14:17 doi:10.1186/1471-2091-14-17