Journal of Inherited Metabolic Disease

, Volume 13, Issue 5, pp 692–700 | Cite as

Identification of a single nucleotide substitution in the coding sequence ofin vitro amplified cDNA from a patient with partial HPRT deficiency (HPRTBrisbane)

  • R. B. Gordon
  • D. G. Sculley
  • P. A. Dawson
  • I. R. Beacham
  • B. T. Emmerson


The change in genomic DNA responsible for HPRT deficiency has been determined in a patient with urate overproduction and gout. In erythrocyte cell lysates, this patient had approximately 10% of normal HPRT enzyme activity and 26% of immunoidentical HPRT protein. Cultured lymphoblasts derived from this patient were used to extract mRNA. This was reverse transcribed to cDNA, which was then amplified using the polymerase chain reaction. The resulting DNA was cloned and the nucleotide sequence determined. In addition a portion of the sequence was derived from cloned double-stranded cDNA prepared by conventional first and second strand synthesis. A single nucleotide base change (a C → T transition) was detected, which predicts an amino acid substitution of isoleucine for threonine at amino acid 168 of the HPRT protein. The nucleotide substitution creates aBamHI site, confirming a restriction fragment length polymorphism previously reported in this patient.


Threonine Restriction Fragment Length Polymorphism Gout Fragment Length Polymorphism Amino Acid Substitution 
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Copyright information

© SSIEM and Kluwer Academic Publishers 1990

Authors and Affiliations

  • R. B. Gordon
    • 1
  • D. G. Sculley
    • 1
  • P. A. Dawson
    • 1
  • I. R. Beacham
    • 2
  • B. T. Emmerson
    • 1
  1. 1.Department of MedicineUniversity of Queensland, Princess Alexandra HospitalBrisbaneAustralia
  2. 2.Division of Science and TechnologyGriffith UniversityBrisbaneAustralia

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