The primary transcription unit of the human (eqn)a2 globin gene defined by quantitative RT/PCR

Abstract

We have set up an experimental system to map the primary transcription unit of the human α2 globin gene. The duplicated human α globin genes (α2-α1) were linked to the α globin locus Positive Regulatory Element (PRE) and stably transfected into murine erythroleukaemia cells. We then developed a quantitative reverse transcriptase, polymerase chain reaction assay to map α2 primary transcripts using primer pairs derived from different parts of the α2 globin gene and its 3′ flanking region. This approach has revealed the presence of steady state nuclear RNA past the poly(A) site of the α2 globin gene at approximately 40% of the level of unspliced Intron transcript. Furthermore, these 3′ flanking transcripts diminish 500 bp into the 3′ flanking region, identifying this part of the α2 globin gene as the principal region of termination of transcription.