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Abstract
The results presented in this communication demonstrate that hypoxanthine-guanine phosphoribosyltransferase (HPRT) cDNA can be expressed in both Chinese hamster and human fibroblasts deficient in the endogenous gene product at levels permitting normal growth of the transformants. All the elements necessary for this expression are present in a pBR322-derived plasmid containing HPRT cDNA coding sequence and a retroviral long terminal repeat. These molecules function in both species investigated and, at least in the case of the Chinese hamster transformants, are efficient at the single copy level. Although the effects of the presence of intron sequences and a polyadenylation signal within the plasmids have yet to be evaluated, these studies demonstrate that neither is an absolute requirement for expression of HPRT cDNA sequences in cultured mammalian cells. We describe the construction of recombinant plasmids containing wild type human or Chinese hamster HPRT cDNA sequences in tandem with a retroviral LTR which confer the HPRT+ phenotype in HPRT-deficient V79 and Lesch-Nyhan fibroblasts. Both stable and unstable transformants, that expressed HPRT mRNA and protein, were isolated at high frequency.
Highlights
Recombinants in Cultured LeschNyhan and Chinese Hamster acterization of murine H P R T cDNA sequences [5,6] isolated froma mouse neuroblastoma cell line that overproduceda mutant HPRT proteibny virtue of amplification of the HPRT
These sequenceshavebeen used as probes to identify andisolate H P R T cDNA sequences from a (Received for publication, April 28, 1983) wild type Chinese hamsterV79 cell line RJKO [9] and from a
We describe the construction of recombinant plasmids containing wildtype human or Chinese hamster HPRTcDNA sequences in tandem with a retroviral LTR which con
Summary
Recombinants in Cultured LeschNyhan and Chinese Hamster acterization of murine H P R T cDNA sequences [5,6] isolated froma mouse neuroblastoma cell line that overproduceda mutant HPRT proteibny virtue of amplification of the HPRT. These sequenceshavebeen used as probes to identify andisolate H P R T cDNA sequences from a (Received for publication, April 28, 1983) wild type Chinese hamsterV79 cell line RJKO [9] and from a Derived plasmid containing HPRT cDNA coding sequence and a retroviral long terminal repeat.
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