Cloning of human purine-nucleoside phosphorylase cDNA sequences by complementation in Escherichia coli.
Open Access
- 1 July 1983
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 80 (14) , 4281-4285
- https://doi.org/10.1073/pnas.80.14.4281
Abstract
Complementary DNA cDNA clones that contain the entire coding region of the human purine-nucleoside phosphorylase (PNP; EC 2.4.2.1) mRNA were generated by reverse transcription of PNP-enriched mRNA obtained by immunoadsorption of HeLa cell polyribosomes with monospecific antibody to human PNP. cDNA molecules that were close in length to PNP mRNA were separated by agarose gel electrophoresis and inserted into the Pst I site of the plasmid pBR322. Plasmid DNA from the pooled clones was used to transform PNP-deficient E. coli cells; those transformants that phenotypically expressed PNP were isolated on selective media. The presence of human PNP in the selected bacterial cell was detected by immunoprecipitation with human PNP antibody.This publication has 35 references indexed in Scilit:
- Glucocorticoid regulation of protein processing and compartmentalizationNature, 1982
- Nucleotide sequence and evolution of a mammalian α-Tubulin messenger RNAJournal of Molecular Biology, 1981
- Transcriptional control in the production of liver-specific mRNAsCell, 1981
- Analysis of gene control signals by DNA fusion and cloning in Escherichia coliJournal of Molecular Biology, 1980
- Phenotypic expression in E. coli of a DNA sequence coding for mouse dihydrofolate reductaseNature, 1978
- Deoxyguanosine toxicity in a mouse T lymphoma: Relationship to purine nucleoside phosphorylase-associated immune dysfunctionCell, 1978
- Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase IJournal of Molecular Biology, 1977
- NUCLEOSIDE-PHOSPHORYLASE DEFICIENCY IN A CHILD WITH SEVERELY DEFECTIVE T-CELL IMMUNITY AND NORMAL B-CELL IMMUNITYThe Lancet, 1975
- Inherited variants of human nucleoside phosphorylaseAnnals of Human Genetics, 1971
- Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4Nature, 1970