What question did this study set out to answer?

To clone and characterize a full-length cDNA for human hypoxanthine phosphoribosyltransferase (HPRT).

January 1, 1983Open Access

Isolation and characterization of a full-length expressible cDNA for human hypoxanthine phosphoribosyl transferase.

Key Points

To clone and characterize a full-length cDNA for human hypoxanthine phosphoribosyltransferase (HPRT).
Cloning a 1.6-kilobase cDNA of human HPRT into a simian virus 40-based expression vector.
Determining the full nucleotide sequence of the cloned cDNA.
Transfecting HPRT-deficient mouse LA9 cells with the purified plasmid to assess enzyme activity.
The inferred amino acid sequence from the cDNA matches a previously identified partial sequence of authentic human HPRT protein.
Transfected LA9 cells successfully expressed human HPRT enzyme activity after selection in hypoxanthine/aminopterin/thymidine medium.

Abstract

We have cloned a full-length 1.6-kilobase cDNA of a human mRNA coding for hypoxanthine phosphoribosyltransferase (HPRT; IMP:pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) into a simian virus 40-based expression vector and have determined its full nucleotide sequence. The inferred amino acid sequence agrees with a partial amino acid sequence determined for authentic human HPRT protein. Transfection of HPRT-deficient mouse LA9 cells with the purified plasmid leads to the expression of human HPRT enzyme activity in cells stably transfected and selected for enzyme activity in hypoxanthine/aminopterin/thymidine medium.

Isolation and characterization of a full-length expressible cDNA for human hypoxanthine phosphoribosyl transferase.

Key Points

Abstract

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