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Biomedical subjects

F H Ruddle

Publications and source records attributed to F H Ruddle.

At least 217 records · Page 12Linked to original sources
PubMed1982

Genetic and biochemical characterization of a human surface determinant on somatic cell hybrids: the 4F2 antigen.

We have mapped the gene which codes the species-specific determinant defined by monoclonal antibody 4F2 to human chromosome 11. All human chromosomes, except Y, were included in a group of four human-mouse hybrid lines. Hybrids heterogeneous for 4F2 antigen expression were sorted using the fluorescence-activated cell sorter (FACS) to yield populations homogeneous with respect to the presence or absence of this determinant. Isozyme analysis indicated corresponding genetic selection for or against human chromosome 11. This map assignment was confirmed using a hybrid line which contained only human chromosome 11. Immunoprecipitation of the 4F2 determinant from the 11 only hybrid resulted in a heavy subunit of molecular weight (Mr) = 100,000 and a light subunit of Mr = 41,000. This contrasts with results obtained from nonhybrid human cells of different lineages. These results demonstrate the importance of FACS techniques in the rapid mapping of genes which code human cell surface antigens.

Animals↗
PubMed1982

Dispersion of argininosuccinate-synthetase-like human genes to multiple autosomes and the X chromosome.

DNA sequences closely homologous to argininosuccinate synthetase are present at ten or more distinct locations in the human genome, including sites on chromosomes 6, 9 and X. Argininosuccinate synthetase thus represents one of the most widely dispersed multigene families described to date, the first instance of a multigene family associated with an enzyme of intermediary metabolism and, perhaps most striking, the first instance of a multigene family with members on both autosomes and sex chromosomes.

Argininosuccinate Synthase↗
PubMed1982

Partial purification and characterization of the mRNA for human thymidine kinase and hypoxanthine/guanine phosphoribosyltransferase.

We used direct microinjection of poly(A)+RNA into individual hypoxanthine/guanine phosphoribosyltransferase-deficient or thymidine kinase-deficient cells and detected the specific in vivo translation products as an assay for human hypoxanthine/guanine phosphoribosyltransferase or thymidine kinase mRNAs. The incorporation of [3H]hypoxanthine or [3H]thymidine into cells in response to injected mRNA was assayed in situ by autoradiography. Methylmercuric hydroxide/agarose gel analysis showed that human hypoxanthine/guanine phosphoribosyltransferase mRNA contains approximately 1,530 nucleotides, which is twice the number required for its protein coding capacity. The mRNA for human cytoplasmic thymidine kinase is estimated to be approximately the same length; thus, the size of the cytosol thymidine kinase subunit can be predicted to be approximately 47,000 daltons, if the full coding capacity of its mRNA is utilized.

Biological Assay↗
PubMed1982

Mechanisms of DNA uptake by mammalian cells: fate of exogenously added DNA monitored by the use of fluorescent dyes.

Coprecipitation of DNA with calcium phosphate is a commonly used method of gene transfer in mammalian cells. We have found that DNA forms a tight complex with Ca Pi and that DNA in this complex is resistant to nucleases present in serum or added externally. Under optimal conditions, virtually all of the recipient mouse Ltk- Aprt- cells take up Ca Pi--DNA complexes, as determined by fluorescent dyes specific for DNA (4',6-diaminilo-2-phenylindol dihydrochloride) or for calcium salts (chlorotetracycline). However, only a small proportion of the cells have detectable CA Pi--DNA complexes in the nucleus. Uptake of the Ca Pi--DNA complexes was highly dependent upon the pH at which the Ca Pi--DNA complexes was formed and upon the concentration of DNA in the complex.

Animals↗
PubMed1982

Identification of human genomic clones coding the major histocompatibility antigens HLA-a2 and HLA-B7 by DNA-mediated gene transfer.

We have screened a large number of isolated human genomic clones that hybridize to a cloned HLA cDNA probe for their ability to direct the synthesis of HLA-A, -B, and -C surface antigens on mouse L cells following DNA-mediated gene transfer. The surface expression of human histocompatibility antigens, monitored by indirect immunofluorescence and the fluorescence-activated cell sorter, was examined at 60 hr after transfection and on hypoxanthine/aminopterin/thymidine-resistant (HATR) populations derived from cotransfer with the herpes simplex virus thymidine kinase gene. Two unique genomic clones designated JY B3.2 and JY 158, isolated from the human lymphoblastoid cell line JY (homozygous HLA-A2, -B7), were shown to contain gene sequences capable of directing expression of an HLA-A, -B, -C determinant. By using allo-specific antibodies, the gene products of these clones were identified as HLA-A2 and HLA-B7, respectively. HATR clonal populations isolated from cotransfections with these genomic clones displayed varying levels of surface HLA expression that correlated with the number of intact donor HLA sequences present in the cells. In general, these levels of expression were stable during 3 months in culture. This system provides a powerful tool for the study of human surface antigen gene structure, expression, and function on a mouse cell background.

Antigens, Surface↗
PubMed1982

DNA sequence associated with chromosome translocations in mouse plasmacytomas.

A DNA sequence that generates aberrantly rearranged immunoglobulin heavy chain constant region genes in murine plasmacytomas is shown to participate in a chromosome translocation. We have previously termed this DNA sequence NIARD for non-immunoglobulin-associated rearranging DNA. NIARD rearrangements were found frequently in murine plasmacytomas but were not detected in normal lymphocytes. These rearrangements occasionally involve the switch region of the C alpha gene. In this study, DNA samples obtained from mouse-Chinese hamster somatic cell hybrid lines were digested with various restriction endonucleases and analyzed by the Southern transfer technique with a NIARD hybridization probe. These experiments show that NIARD resides on chromosome 15 in the mouse germ line. Since NIARD is found adjacent to the C alpha gene (located on chromosome 12) in some plasmacytomas, it is apparent that a translocation involving these two chromosomes has occurred. We have proposed a rcpT(12;15) model to explain our data. The implications of NIARD rearrangements for malignant transformation are discussed.

Animals↗
PubMed1982

Localization of the casein gene family to a single mouse chromosome.

A series of mouse-hamster somatic cell hybrids containing a variable number of mouse chromosomes and a constant set of hamster chromosomes have been used to determine the chromosomal location of a family of hormone-inducible genes, the murine caseins. Recombinant mouse cDNA clones encoding the alpha-, beta-, and gamma-caseins were constructed and used in DNA restriction mapping experiments. All three casein cDNAs hybridized to the same set of somatic cell hybrid DNAs isolated from cells containing mouse chromosome 5, while negative hybridization was observed to ten other hybrid DNAs isolated from cells lacking chromosome 5. A fourth cDNA clone, designated pCM delta 40, which hybridized to an abundant 790 nucleotide poly(A)RNA isolated from 6-d lactating mouse mammary tissue, was also mapped to chromosome 5. The chromosomal assignment of the casein gene family was confirmed using a mouse albumin clone. The albumin gene had been previously localized to mouse chromosome 5 by both breeding studies and analogous molecular hybridization experiments. An additional control experiment demonstrated that another hormone-inducible gene, specifying a 620 nucleotide abundant mammary gland mRNA, hybridized to DNA isolated from a different somatic cell hybrid line. These studies represent the first localization of a peptide and steroid hormone-responsive gene family to a single mouse chromosome.

Animals↗
PubMed1982

Chromosomal localization of the Moloney sarcoma virus mouse cellular (c-mos) sequence.

The Moloney sarcoma virus-specific onc gene, referred to as v-mos, was used as probe to hybridize to restricted DNAs from various mouse-Chinese hamster hybrid cell lines. These hybrid cells contain, in addition to all of the Chinese hamster chromosomes, various numbers (less than a full complement) of mouse chromosomes. Comparison of the presence or absence of the mouse cellular mos gene with the known karyotype in each of the hybrid cell lines allows us to conclude that the mos gene is on mouse chromosome 4.

Animals↗
PubMed1982

Human beta interferon gene localization and expression in somatic cell hybrids.

The human fibroblast interferon gene beta 1 was mapped to human chromosome 9. Sequence homology with a beta 1 cDNA clone was detected in both genomic DNA and induced mRNA of human/mouse or human/hamster somatic cell hybrids containing human chromosome 9, but not in lines lacking this chromosome or those retaining a complex translocation involving chromosomes 9 and 11. Interferon mRNA that did not share sequence homology with the beta 1 cDNA clone was detected in lines containing human chromosomes 2 and 5 but lacking chromosome 9, suggesting the presence of other unlinked interferon sequences in the human genome.

Animals↗
PubMed1982

Comparative gene mapping: murine lambda light chain genes are located in region cen to B5 of mouse chromosome 16 not homologous to human chromosome 21.

We have mapped the genes for murine immunoglobulin lambda light chains to the region of chromosome 16 proximal to band B5 by hybridizing a cDNA probe for gamma light chains to the DNA of a series of hybrid clones made between mouse fibroblasts carrying Searle's translocation, T(X;16)16H, and Chinese hamster cells. Based on homology, we predict that the human Ig gamma gene (IGL) will map to the proximal two-thirds of HSA 22.

Animals↗
PubMed1982

Germ line transmission in transgenic mice.

Our laboratory has recently demonstrated the successful transfer of genetic material into the genomes of newborn mice by injection of that material into pronuclei of fertilized eggs. Our initial results indicated two patterns of processing of the injected DNA: one in which the material was not integrated into the host genome, and another in which the injected genes became associated with high molecular weight DNA. We now present evidence that these patterns are maintained through further development to adulthood. We provide definitive evidence of the covalent association of injected DNA with host sequences, and transmission of such linked sequences in a Mendelian distribution to 2 succeeding generations of progeny. The observation that injected gene sequences can become integrated and stably transmitted through the germ line should contribute significantly to further studies of this gene transfer system.

Aging↗
PubMed1981

Integration and stable germ line transmission of genes injected into mouse pronuclei.

Genetic material has been successfully transferred into the genomes of newborn mice by injection of that material into pronuclei of fertilized eggs. Initial results indicated two patterns of processing the injected DNA: one in which the material was not integrated into the host genome, and another in which the injected genes became associated with high molecular weight DNA. These patterns are maintained through further development to adulthood. The evidence presented indicates the covalent association of injected DNA with host sequences, and transmission of such linked sequences in a Mendelian distribution to two succeeding generations of progeny.

Animals↗
PubMed1981

A new era in mammalian gene mapping: somatic cell genetics and recombinant DNA methodologies.

Mammalian gene mapping techniques are now sufficiently advanced to contribute significantly to prenatal diagnosis and to human molecular genetics. Restriction fragment mapping can be used to place polymorphic genetic markers at random sites within the genome, and these sites used to assign genes responsible for disease conditions to a chromosomal region. Somatic cell genetic techniques can then be applied to saturate that region with additional restriction fragment markers, some of which will be closely linked to the disease gene. Closely linked restriction fragment markers, especially flanking pairs of markers, can act as predictors for the transmission of defective genes to offspring. A series of tightly linked flanking restriction markers might in addition contribute to the eventual isolation and cloning of the disease gene itself.

Animals↗