Isolation of a Genomic Clone Partially Encoding Human Hypoxanthine Phosphoribosyltransferase

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1982 Aug 1

PMID 6956912

Citations 43

Authors

D J Jolly

A C Esty

H U Bernard

T Friedmann

Affiliations

Soon will be listed here.

Abstract

Mouse cells deficient in the enzyme hypoxanthine phosphoribosyltransferase (HPRT; EC 2.4.2.8) have been transfected with total human DNA, and cells producing human enzyme were isolated by growth in selective medium. DNA from several such cell lines has been used to generate secondary transfectants that make human HPRT. Blots of the DNA of these secondary cells have been hybridized with total human DNA probes or with cloned human Alu sequences, and one of several common bands has been cloned in pBR322. Colonies of transformed Escherichia coli containing human sequences were detected by their homology with human DNA, and subclones of resulting recombinant plasmids were prepared. Two subclones free of Alu sequences were found to contain human sequences that hybridized to human X chromosome DNA. One of these, pBR1.5, also hybridized to a single RNA band on gel blots of human and secondary transfectant cytoplasmic poly(A)+RNA but not to RNA from the parent mouse cell line. These results indicate that these clones represent human HPRT gene fragments. This has been confirmed by using pBR1.5 as a probe to isolate an authentic and expressible human HPRT cDNA clone from a library prepared by H. Okayama and P. Berg.

Citing Articles

Extending AAV Packaging Cargo through Dual Co-Transduction: Efficient Protein Trans-Splicing at Low Vector Doses.

Ferreira M, Fernandes S, Almeida A, Neto S, Mendes J, Silva R Int J Mol Sci. 2023; 24(13).

PMID: 37445701 PMC: 10341399. DOI: 10.3390/ijms241310524.

Functional cDNA expression cloning: pushing it to the limit.

Okayama H Proc Jpn Acad Ser B Phys Biol Sci. 2012; 88(3):102-19.

PMID: 22450538 PMC: 3365248. DOI: 10.2183/pjab.88.102.

A bovine papilloma virus vector with a dominant resistance marker replicates extrachromosomally in mouse and E. coli cells.

Matthias P, Bernard H, Scott A, Brady G, Schutz G EMBO J. 1983; 2(9):1487-92.

PMID: 11892800 PMC: 555311. DOI: 10.1002/j.1460-2075.1983.tb01612.x.

DNA replication analysis of FMR1, XIST, and factor 8C loci by FISH shows nontranscribed X-linked genes replicate late.

Torchia B, Call L, MIGEON B Am J Hum Genet. 1994; 55(1):96-104.

PMID: 8023856 PMC: 1918226.

Human hypoxanthine-guanine phosphoribosyltransferase: studies of the normal and five mutant forms of the enzyme.

Kelley W, Wilson J Trans Am Clin Climatol Assoc. 1982; 94:91-9.

PMID: 7186238 PMC: 2279576.

References

Becker M, Seegmiller J . Recent advances in the identification of enzyme abnormalities underlying excessive purine synthesis in man. Arthritis Rheum. 1975; 18(6 Suppl):687-94. DOI: 10.1002/art.1780180708. View

Deininger P, Schmid C . An electron microscope study of the DNA sequence organization of the human genome. J Mol Biol. 1976; 106(3):773-90. DOI: 10.1016/0022-2836(76)90264-3. View

Ullrich A, Shine J, Chirgwin J, Pictet R, Tischer E, Rutter W . Rat insulin genes: construction of plasmids containing the coding sequences. Science. 1977; 196(4296):1313-9. DOI: 10.1126/science.325648. View

Cory S, Adams J . A very large repeating unit of mouse DNA containing the 18S, 28S and 5.8S rRNA genes. Cell. 1977; 11(4):795-805. DOI: 10.1016/0092-8674(77)90292-6. View

Bolivar F, Rodriguez R, Greene P, Betlach M, Heyneker H, Boyer H . Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977; 2(2):95-113. View

Graham D . The isolation of high molecular weight DNA from whole organisms or large tissue masses. Anal Biochem. 1978; 85(2):609-13. DOI: 10.1016/0003-2697(78)90262-2. View

Chang A, Cohen S . Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978; 134(3):1141-56. PMC: 222365. DOI: 10.1128/jb.134.3.1141-1156.1978. View

Levis R, Penman S . Processing steps and methylation in the formation of the ribosomal RNA of cultured Drosophila cells. J Mol Biol. 1978; 121(2):219-38. DOI: 10.1016/s0022-2836(78)80006-0. View

Pellicer A, Wigler M, Axel R, Silverstein S . The transfer and stable integration of the HSV thymidine kinase gene into mouse cells. Cell. 1978; 14(1):133-41. DOI: 10.1016/0092-8674(78)90308-2. View

10.

Wigler M, Pellicer A, Silverstein S, Axel R . Biochemical transfer of single-copy eucaryotic genes using total cellular DNA as donor. Cell. 1978; 14(3):725-31. DOI: 10.1016/0092-8674(78)90254-4. View

11.

Philippsen P, Kramer R, Davis R . Cloning of the yeast ribosomal DNA repeat unit in SstI and HindIII lambda vectors using genetic and physical size selections. J Mol Biol. 1978; 123(3):371-86. DOI: 10.1016/0022-2836(78)90085-2. View

12.

Lehrach H, Frischauf A, Hanahan D, Wozney J, Fuller F, Crkvenjakov R . Construction and characterization of a 2.5-kilobase procollagen clone. Proc Natl Acad Sci U S A. 1978; 75(11):5417-21. PMC: 392975. DOI: 10.1073/pnas.75.11.5417. View

13.

Johnson G, Eisenberg L, MIGEON B . Human and mouse hypoxanthine-guanine phosphoribosyltransferase: dimers and tetramers. Science. 1979; 203(4376):174-6. DOI: 10.1126/science.569362. View

14.

Caskey C, Kruh G . The HPRT locus. Cell. 1979; 16(1):1-9. DOI: 10.1016/0092-8674(79)90182-x. View

15.

Becker M, Yen R, ITKIN P, Goss S, Seegmiller J, BAKAY B . Regional localization of the gene for human phosphoribosylpyrophosphate synthetase on the X chromosome. Science. 1979; 203(4384):1016-9. DOI: 10.1126/science.218284. View

16.

Smith A . The use of exonuclease III for preparing single stranded DNA for use as a template in the chain terminator sequencing method. Nucleic Acids Res. 1979; 6(3):831-48. PMC: 327737. DOI: 10.1093/nar/6.3.831. View

17.

Wahl G, Stern M, Stark G . Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979; 76(8):3683-7. PMC: 383897. DOI: 10.1073/pnas.76.8.3683. View

18.

Duncan C, Biro P, Choudary P, Elder J, Wang R, Forget B . RNA polymerase III transcriptional units are interspersed among human non-alpha-globin genes. Proc Natl Acad Sci U S A. 1979; 76(10):5095-9. PMC: 413086. DOI: 10.1073/pnas.76.10.5095. View

19.

Gergen J, STERN R, Wensink P . Filter replicas and permanent collections of recombinant DNA plasmids. Nucleic Acids Res. 1979; 7(8):2115-36. PMC: 342374. DOI: 10.1093/nar/7.8.2115. View

20.

Peterson J, MCBRIDE O . Cotransfer of linked eukaryotic genes and efficient transfer of hypoxanthine phosphoribosyltransferase by DNA-mediated gene transfer. Proc Natl Acad Sci U S A. 1980; 77(3):1583-7. PMC: 348541. DOI: 10.1073/pnas.77.3.1583. View