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HGPRTHypoxanthine-Guanine Phosphoribosyl Transferase
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The following primers were used: HGPRT (362 bp) sense 5'-GTT GGA TAC AGG CCA GAC TTT GTT G-3', antisense 5'-GAG GGT AGG CTG GCC TAT AGG CT-3'; interleukin (IL)-1 (400 bp) sense 5'-GCA ACT GTT CCT GAA CTC A-3', antisense 5'-CTC GGA GCC TGT AGT GCA G-3'; IL-10 (256 bp) sense 5'-TAC CTG GTA GAA GTG ATG CC-3', antisense 5'-CAT CAT GTA TGC TTC TAT GC-3'; IL-12 (312 bp) sense 5'-CGT GCT CAT GGC TGG TGC AAA G-3', antisense 5'-CTT CAT CTG CAA GTT CTT GGG C-3'; IL-18 (440 bp) sense 5'-ACT GTA CAA CCG GAG TAA TAC GG-3', antisense 5'-AGT GAA CAT TAC AGA TTT ATC CC-3'; tumour necrosis factor (TNF)-[alpha] (276 bp) sense 5'-ATG AGC ACA GAA AGC ATG ATC-3', antisense 5'-TAC AGG CTT GTC ACT CGA ATT-3'; IFN[.
A proton nuclear magnetic resonance (1H-NMR) spectrometric method can be used to measure many compounds of purine and pyrimidine metabolism (13), but it has a major disadvantage in that it fails to detect uric acid and 2,8-dihydroxyadenine, which are very useful markers for the diagnosis of APRT deficiency, XDH deficiency, molybdenum cofactor deficiency, PRPPS super-activity, and HGPRT deficiency.
HGPRT catalyzes a biochemical process called purine salvage.
After an initial denaturation for 69s at 95 [degrees], 36 cycles of amplification (93[degrees]C for 55s, 61[degrees]C for 45s, 72 [degrees]C for 40s) for TNF-[alpha] and HGPRT (hypoxanthine-guanine-phosphoribosyltransferase, as a positive control and internal standard) cDNAs were performed followed by a 100-s step for elongation at 72 [degrees].