Real-time quantitative RT-PCR for human telomere elongation reverse transcriptase in chronic myeloid leukemia.
Telomeres cap chromosome ends and are pivotal for DNA stability. Deregulation of the telomere stabilising enzyme telomerase in malignancy has implications in diagnosis, prognosis and therapeutics of cancer. Quantification of the expression of the telomerase catalytic subunit, hTERT, using the LightCycler TeloTAGGG hTERT Quantification kit is not optimal for analysis of chronic myeloid leukemia (CML) samples. The internal control, porphobilinogen deaminase (PBGD) is amplified in a separate tube to hTERT and has an unstable genomic localisation of 11q23. Our laboratory thus developed a real-time reverse transcriptase polymerase chain reaction which co-amplifies hTERT and either mitochondrial single-stranded DNA binding protein 1 (ssBP1) or ubiquitin C (UBC).
Gil
Marcel E
ME
Department of Molecular Medicine and Haematology, School of Pathology, University of the Witwatersrand, 7 York Road, Parktown, 2193 Johannesburg, South Africa. marcelg@mweb.co.za
Coetzer
Thérèsa L
TL
D004268:DNA-Binding Proteins / Q000592:standards; D006801:Humans; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000201:enzymology* / Q000235:genetics; D012334:RNA, Neoplasm / Q000032:analysis*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D019098:Telomerase / Q000032:analysis / Q000235:genetics*; D025822:Ubiquitin C / Q000592:standards
D004268:DNA-Binding Proteins; D012334:RNA, Neoplasm; D025822:Ubiquitin C; D019098:Telomerase