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Literature DB >> 3889850

Rapid assay for detection of Escherichia coli xanthine-guanine phosphoribosyltransferase activity in transduced cells.

Abstract

Cultured mammalian cells transduced with the Escherichia coli gene, Ecogpt, synthesize the bacterial enzyme xanthine-guanine phosphoribosyl transferase (XGPT) (1). This paper describes a method for measuring XGPT activity in crude cell extracts by following the conversion of 14C-xanthine (X) to 14C-xanthine monophosphate (XMP) and 14C-xanthosine (XR) by thin layer chromatography. The method is rapid, easy to use, sensitive and linear over a wide range of XGPT activity and has been useful for detecting XGPT in cells that were transiently transfected or stably transformed with Ecogpt. During our studies, we have found that a human cell line (XP20S) converts xanthine to XMP. This activity is probably catalyzed by a variant hypoxanthine-guanine phosphoribosyltransferase (HGPT) since the low activity is readily inhibited by hypoxanthine. A low level of conversion of X to XMP may explain why some cell lines are not killed in a medium containing mycophenolic acid and X.

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Year: 1985 PMID： 3889850 PMCID： PMC341204 DOI： 10.1093/nar/13.8.2921

Source DB: PubMed Journal: Nucleic Acids Res ISSN： 0305-1048 Impact factor: 16.971

16 in total

1. Guanine phosphoribosyltransferase from Escherichia coli, specificity and properties.

Authors: R L Miller; G A Ramsey; T A Krenitsky; G B Elion
Journal: Biochemistry Date: 1972-12-05 Impact factor: 3.162

2. A new technique for the assay of infectivity of human adenovirus 5 DNA.

Authors: F L Graham; A J van der Eb
Journal: Virology Date: 1973-04 Impact factor: 3.616

3. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells.

Authors: R Radloff; W Bauer; J Vinograd
Journal: Proc Natl Acad Sci U S A Date: 1967-05 Impact factor: 11.205

4. Mapping temperature-sensitive and host-range mutations of adenovirus type 5 by marker rescue.

Authors: E Frost; J Williams
Journal: Virology Date: 1978-11 Impact factor: 3.616

5. Regulation of simian virus 40 transcription: sensitive analysis of the RNA species present early in infections by virus or viral DNA.

Authors: B A Parker; G R Stark
Journal: J Virol Date: 1979-08 Impact factor: 5.103

6. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex.

Authors: L Katz; D T Kingsbury; D R Helinski
Journal: J Bacteriol Date: 1973-05 Impact factor: 3.490

7. Comparative studies of host-cell reactivation, colony forming ability and excision repair after UV irradiation of xeroderma pigmentosum, normal human and some other mammalian cells.

Authors: H Takebe; S Nii; M I Ishii; H Utsumi
Journal: Mutat Res Date: 1974-12 Impact factor: 2.433

8. Lesch-Nyhan syndrome: altered kinetic properties of mutant enzyme.

Authors: J A McDonald; W N Kelley
Journal: Science Date: 1971-02-19 Impact factor: 47.728

9. Purification and characterization of Escherichia coli guanine-xanthine phosphoribosyltransferase produced by a high efficiency expression plasmid utilizing a lambda PL promoter and CI857 temperature-sensitive repressor.

Authors: S W Liu; G Milman
Journal: J Biol Chem Date: 1983-06-25 Impact factor: 5.157

10. The inhibition of nucleic acid synthesis by mycophenolic acid.

Authors: T J Franklin; J M Cook
Journal: Biochem J Date: 1969-07 Impact factor: 3.857

15 in total

1. The nucleotide sequence of greA, a suppressor gene that restores growth of an Escherichia coli RNA polymerase mutant at high temperature.

Authors: J Sparkowski; A Das
Journal: Nucleic Acids Res Date: 1990-11-11 Impact factor: 16.971

2. Mouse embryonic stem cells exhibit high levels of extrachromosomal homologous recombination in a chloramphenicol acetyltransferase assay system.

Authors: M Jasin; F Liang
Journal: Nucleic Acids Res Date: 1991-12 Impact factor: 16.971

Rapid assay for detection of Escherichia coli xanthine-guanine phosphoribosyltransferase activity in transduced cells.

1. Guanine phosphoribosyltransferase from Escherichia coli, specificity and properties.

2. A new technique for the assay of infectivity of human adenovirus 5 DNA.

3. A dye-buoyant-density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HeLa cells.

4. Mapping temperature-sensitive and host-range mutations of adenovirus type 5 by marker rescue.

5. Regulation of simian virus 40 transcription: sensitive analysis of the RNA species present early in infections by virus or viral DNA.

6. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex.

7. Comparative studies of host-cell reactivation, colony forming ability and excision repair after UV irradiation of xeroderma pigmentosum, normal human and some other mammalian cells.

8. Lesch-Nyhan syndrome: altered kinetic properties of mutant enzyme.

9. Purification and characterization of Escherichia coli guanine-xanthine phosphoribosyltransferase produced by a high efficiency expression plasmid utilizing a lambda PL promoter and CI857 temperature-sensitive repressor.

10. The inhibition of nucleic acid synthesis by mycophenolic acid.

1. The nucleotide sequence of greA, a suppressor gene that restores growth of an Escherichia coli RNA polymerase mutant at high temperature.

2. Mouse embryonic stem cells exhibit high levels of extrachromosomal homologous recombination in a chloramphenicol acetyltransferase assay system.

3. Deletion of stably integrated DNA is suppressed by cadmium and zinc.

4. The cell-specific elastase I enhancer comprises two domains.

5. Electroporation for the efficient transfection of mammalian cells with DNA.

6. Germline sequence of two human T-cell receptor V beta genes: V beta 8.1 is transcribed from a TATA-box promoter.

7. A rapid and sensitive assay system for bacterial gpt activity in transfected mammalian cells.

8. Fast non-chromatographic method to assay for xanthine-guanosine phosphoribosyl transferase.

9. High-efficiency transformation of bacterial cells by electroporation.

10. Cisplatin-resistant cells express increased levels of a factor that recognizes damaged DNA.