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Biomedical subjects

F H Ruddle

Publications and source records attributed to F H Ruddle.

At least 289 records · Page 16Linked to original sources

Genetic analysis of the human cell surface: antigenic marker for the human X chromosome in human-mouse hybrids.

Somatic cell hybrids of human fibroblasts and mouse A9 cells, carrying only a portion of the human X chromosome in a mouse chromosome background, were injected into C3H mice. The resulting mouse anti-hybrid cell antisera contain antibodies found to be human specific and to react with only those hybrid cells carrying the human X chromosome, as confirmed by essentially perfect concordance between antibody binding assayed by indirect immunofluorescence and presence of the human X-linked enzyme hypoxanthine phosphoribosyl-transferase determined by autoradiographic assay of [3H] hypoxanthine utilization. Heterogeneous mixtures of hybrid cells may be analyzed into fluorescent (X plus) and nonfluorescent (X minus) subpopulations and fractionated viably by using a fluorescence-activated cell sorter.

Animals↗

Co-transfer of human X-linked markers into murine somatic cells via isolated metaphase chromosomes.

Transformation frequencies of 4 x 10(-5) were obtained in chromosome-mediated gene transfer experiments using human cell line HeLa S3 as donor and mouse cell line A9 as recipient. This high frequency of interspecific transformation was achieved by treating the recipient cells with dimethylsulfoxide in addition to other facilitators. The high frequency of transformation correlated positively with transgenome size on the basis of both co-transfer of linked markers and chromosome analysis. The syntenic human markers glucose-6-phosphate dehydrogenase (D-glucose-6-phosphate:NADP(+) 1-oxidoreductase, EC 1.1.1.49) and phosphoglycerate kinase (ATP:3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3) were sometimes transferred together with the selected X-linked prototrophic marker hypoxanthine phosphoribosyltransferase (IMP: pyrophosphate phosphoribosyltransferase, EC 2.4.2.8) into murine somatic cells. Donor human chromosome material could be demonstrated cytologically in some of the transformed cell lines. Transformants exhibited various rates of loss of the human hypoxanthine phosphoribosyltransferase marker when grown under nonselective conditions. These results reveal a broader range of possible interspecific transgenome sizes than has been recognized in the past. The largest transgenomes consist of cytologically detectable donor fragments and contain syntenic markers that are not closely linked to the selected marker.

Dimethyl Sulfoxide↗

Presence of human chromosome 21 alone is sufficient for hybrid cell sensitivity to human interferon.

Human/mouse somatic cell hybrids with chromosome 21 as the only detectable human genetic material were sensitive to both human leukocyte and fibroblast interferons. The presence of additional human chromosomes decreased the amount of interferon needed to attain a given level of virus resistance. Decreased cytopathic effects, decreased virus yields, and the appearance of a specific phosphorylated protein associated with interferon treatment were all observed in hybrids maintaining only human chromosome 21. The phosphorylated protein found in extracts of these human interferon-treated hybrid cells was of mouse origin.

Animals↗

Assignment of the gene for adenosine kinase to chromosome 14 in Mus musculus by somatic cell hybridization.

Evidence is presented for the assignment of the gene for adenosine kinase to Mus musculus chromosome 14 by synteny testing and karyotypic analysis of mouse X Chinese hamster somatic cell hybrid clones. ADOK and two enzymes previously mapped to mouse chromosome 14, NP and ES-10, were expressed concordantly in 29 hybrid clones. Chromosome analysis confirmed this assignment. Syntenic evidence is also presented using several clones of a gene transfer system in which the gene for human HPRT had integrated into modified mouse chromosome 14's.

Adenosine Kinase↗

Stimulation of in vitro translation of messenger RNA by actinomycin D and cordycepin.

Actinomycin D and cordycepin were tested for their effect on translation in the wheat germ embryo extract and reticulocyte lysate assays for in vitro protein synthesis. Both drugs were found to stimulate the incorporation of 35S-labeled methionine into protein up to threefold as compared to control assays. This was true for synthesis directed by murine myeloma polyadenylate-containing RNA, tobacco mosaic virus RNA, and endogenous reticulocyte messenger RNA.

Cell-Free System↗

The status of the gene map of the human chromosomes.

In man, the specific chromosome that carries each of about 210 gene loci is known. These loci include at least one assigned to each chromosome (including the Y), about 110 assigned to specific autosomes, and about 100 to the X chromosome. For many loci, information on regional chromosomal localization is also available. The information comes mainly from studies in families and somatic cell hybrids, as well as an intgratsight of results from the two methods. Knowledge of the chromosome map gives insight into evolution, chromosomal organization in relation to genetic control mechanisms, and the pathogenesis of neoplasms and malformations. Furthermore, it is useful in prenatal or premorbid diagnosis of hereditary diseases.

Alleles↗