Levels of hypoxanthine phosphoribosyltransferase RNA in human cells

Journal Article · Thu Feb 01 04:00:00 EST 1990 · Experimental Cell Research; (United States)

DOI:https://doi.org/10.1016/0014-4827(90)90301-P· OSTI ID:5268959

Steen, A M; Luthman, H; Hellgren, D; Lambert, B ^[1]

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1. Karolinska Inst., Stockholm (Sweden)

The gene for the purine salvage enzyme hypoxanthine phosphoribosyltransferase (HPRT) is expressed at a low level in many cells. As is the case with several other housekeeping genes,' thorough studies of hprt gene regulation have been hampered by the low levels of its mRNA. The authors have used RNA/RNA hybridization in solution to determine the concentration of hprt-RNA in human cells. The sensitivity and specificity of the method have been validated, and it is shown that hprt-RNA can be accurately determined at a level of a few mRNA molecules per cell. As expected for a housekeeping gene, low and relatively constant hprt-RNA levels were found in primary cultures of normal amnion cells and fibroblasts, EBV-transformed lymphoblastoid cell lines, neuroblastoma, glioblastoma, and melanoma cell cultures. These results demonstrate the sensitivity of this method for determinations of low levels of RNA and clearly show induction of hprt-RNA after mitogenic stimulation of human lymphocytes.

• OSTI ID: 5268959
• Journal Information: Experimental Cell Research; (United States), Journal Name: Experimental Cell Research; (United States) Vol. 186:2; ISSN ECREA; ISSN 0014-4827
• Country of Publication: United States
• Language: English

Related Subjects

550300* -- Cytology
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
ANIMALS
BIOSYNTHESIS
CONNECTIVE TISSUE CELLS
DISEASES
DNA
ENZYMES
FIBROBLASTS
GENE REGULATION
GLYCOSYL TRANSFERASES
HYPOXANTHINE PHOSPHORIBOSYLTRANSFERASE
MAMMALS
MAN
MELANOMAS
MESSENGER-RNA
MITOGENS
NEOPLASMS
NUCLEIC ACIDS
ORGANIC COMPOUNDS
PENTOSYL TRANSFERASES
PRIMATES
PROTEINS
RECOMBINANT DNA
RNA
SOMATIC CELLS
SYNTHESIS
TRANSFERASES
TUMOR CELLS
VERTEBRATES