Localization of the xanthine guanine phosphoribosyl transferase gene (gpt) of E. coli in AS52 metaphase cells by fluorescence in situ hybridization.
The purpose of this study was to localize the xanthine guanine phosphoribosyl transferase gene (gpt) to a specific chromosome to investigate its proposed autosomal location in the AS52 cell line. AS52 cells are hgprt-deficient Chinese hamster ovary (CHO) cells which carry a single functional copy of the E. coli gpt gene. Fluorescence in situ hybridization (FISH) and digoxigenin-labeled probes, as small as 673 bp, were used in an attempt to localize the 456 bp gpt gene to a specific chromosome. Chi-square analysis of 13 metaphases showed significant labeling on autosomal chromosomes 6 or 7, which are indistinguishable without further banding analysis. Furthermore, a majority of the signals were on the q arm, proximal to the centromere. The data collected supports incorporation of the gpt gene into an acrocentric autosome of the AS52 cell line.
Michaelis
K C
KC
Lilly Research Laboratories, A Division of Eli Lilly and Company, Greenfield, Indiana 46140.
Helvering
L M
LM
Kindig
D E
DE
Garriott
M L
ML
Richardson
K K
KK
D000818:Animals; D001483:Base Sequence; D016466:CHO Cells / Q000166:cytology / Q000378:metabolism; D002896:Chromosomes, Human, Pair 6 / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D002897:Chromosomes, Human, Pair 7 / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D002999:Clone Cells; D006224:Cricetinae; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D016650:Fluorescein-5-isothiocyanate; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence; D008677:Metaphase / Q000235:genetics*; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D010430:Pentosyltransferases / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction
D017931:DNA Primers; D015342:DNA Probes; D010430:Pentosyltransferases; C024827:xanthine phosphoribosyltransferase; D016650:Fluorescein-5-isothiocyanate