Cloning and characterization of 5'-upstream region of human phospholipase C-β2 gene

Abstract

5'-upstream region of the phospholipase C-β2 gene, 810 bp, was cloned and characterized. S1 nuclease mapping and primer extension analyses revealed that a single transcriptional start site locates at 284 nucleotides upstream from the beginning of translation. The 5'-upstream region lacks both TATA motif and typical initiator sequence, but retains GC-rich segment. Two putative regulatory regions, a negative region (−636/−588) and a positive region (−98/ −13) were identified in the upstream region of PLC-β2 gene. We suggest that the transcription of PLC-β2 may be regulated by binding of regulatory proteins to the negative and/or positive regulatory regions located in the upstream of the gene.

Genomic organization and expression of parkin in Drosophila melanogaster

“…The identification of putative binding sites was performed using the MATIN-SPECTOR software (Quandt et al, 1995). To determine the transcription initiation site of D. melanogaster parkin, primer extension analysis was also performed using the Primer Extension System (Promega, Yun et al, 2001). The reverse primer PE was labeled with (γ-33 P) ATP (3,000 Ci/nmol, Amersham), hybridized with RNA and extended by AMV reverse transcriptase.…”