Exon-specific northern analysis and rapid amplification of cDNA ends (RACE) reveal that the proximal promoter II (PII) is responsible for aromatase cytochrome P450 (CYP19) expression in human ovary

摘要: Estrogens are synthesized from C19 steroids by a unique form of cytochrome P450, aromatase P-450 (P-450AROM; the product CYP19 gene). We have shown that tissue-specific expression human P-450AROM is determined, in part, use alternative promoters. Previous methods analysis for determining specific 5'-termini different transcripts included S1 nuclease protection, primer extension, and Northern analysis. In present study we used RACE procedure (rapid amplification cDNA ends) to amplify clone 5' termini expressed corpus luteum (CL). Sequencing resulting clones supports results previously performed studies. Specifically, proximal promoter, PII, predominant promoter utilized CL, such start transcription occurs 26 bp downstream putative TATA sequence. A minority possess an 5'-end, namely I.3. Exon-specific confirms majority CL tissue contain sequence II. Similarly, exon-specific indicates follicles, as well granulosa cells culture, primarily