caa a22 4500 47712657X CHVBK 20180405111651.0 cr unu---uuuuu 170330e19971201xx s 000 0 eng 10.1023/A:1012111325397 doi (NATIONALLICENCE)springer-10.1023/A:1012111325397 Promoter and Intronic Sequences of the Human Thiopurine S-Methyltransferase (TPMT) Gene Isolated from a Human Pacl Genomic Library [Elektronische Daten] [Eugene Krynetski, Michael Fessing, Charles Yates, Daxi Sun, John Schuetz, William Evans] Purpose. To isolate and characterize the polymorphic human thiopurine S-methyltransferase (TPMT) gene. Methods. The human TPMT gene was isolated by PCR screening of a phage artificial chromosome (PAC) library, using exon- and intron-specific primers, then mapped and sequenced. Results. Two separate PAC1 clones were isolated that contained the same 25 kb gene with 9 exons encompassing the entire TPMT open reading frame. Structural characterization revealed distinct differences when compared to a TPMT gene previously isolated from a chromosome 6-specific human genomic library; the 5′-flanking region (putative promoter) contains 17 additional nucleotides located at position-77 upstream from the transcription start site, in addition to several nucleotide sequence differences, and intron 8 is only 1.6 kb, 5 kb shorter than previously reported. Southern and PCR analyses of genomic DNA from 18 unrelated individuals revealed only the TPMT gene structure corresponding to the PAC clones we isolated. Analysis of the TPMT promoter activity using the 5′-terminal region confirmed transcriptional activity in human HepG2 and CCRF-CEM cells. The 5′-flank is 71% GC rich and does not contain consensus sequences for TATA box or CCAAT elements. FISH analysis demonstrated the presence of the TPMT-homologous sequence on the short arm of chromosome 6 (sublocalized to 6p22). Conclusions. These findings establish the genomic structure of the human TPMT gene, revealing differences in the promoter and intronic sequences compared to that previously reported and providing a basis for future studies to further elucidate its biological function and regulation. Plenum Publishing Corporation, 1997 thiopurine S-methyltransferase nationallicence gene structure nationallicence cloning nationallicence Krynetski Eugene Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut Fessing Michael Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut Yates Charles Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut Sun Daxi Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut Schuetz John Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut Evans William Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/12(1997-12-01), 1672-1678 0724-8741 14:12<1672 1997 14 11095 https://doi.org/10.1023/A:1012111325397 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1023/A:1012111325397 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Krynetski Eugene Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut NATIONALLICENCE 700 1- Fessing Michael Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut NATIONALLICENCE 700 1- Yates Charles Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut NATIONALLICENCE 700 1- Sun Daxi Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut NATIONALLICENCE 700 1- Schuetz John Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut NATIONALLICENCE 700 1- Evans William Dept. of Pharmaceutical Sciences, St. Jude Children's Research Hospital, 332 N. Lauderdale St., Memphis, Tennessee 38101 aut NATIONALLICENCE 773 0- Pharmaceutical Research Kluwer Academic Publishers-Plenum Publishers 14/12(1997-12-01), 1672-1678 0724-8741 14:12<1672 1997 14 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer