Report of the Committee on the Genetic Constitution of Chromosomes 18, 19, 20, 21, and 22.
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Biomedical subjects
Publications and source records attributed to A Westerveld.
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A series of man-Chinese hamster somatic cell hybrids with a variable content of human chromosomes was used to study the localization of the human gene coding for the lysosomal enzyme beta-glucocerebrosidase (EC 3.2.1.45). In lysates made from hybrid cells, the human enzyme was specifically recognized by a mouse monoclonal antibody raised against human placental beta-glucocerebrosidase. This monoclonal antibody did not cross-react with Chinese hamster beta-glucocerebrosidase. After reaction of the antibody with the enzyme, beta-glucocerebrosidase was precipitated by addition of Protein A-Sepharose beads, and was detected on the beads by its enzymatic activity. From the analysis of a series of man-Chinese hamster hybrids, among which were hybrids with specific segments of chromosome 1, we conclude that the gene coding for human beta-glucocerebrosidase is localized in the region q21-q31 of chromosome 1.
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In cultured fibroblasts of patients with numerical and structural X chromosome aberrations the activity of steroid sulfatase (STS) is correlated with the number of functional STS gene copies. While normally, this X-linked gene is not inactivated, our data suggest that it may be subject to inactivation when carried on a structurally altered X-chromosome. Similar inactivation patterns have been reported earlier for the Xg locus which, like STS, is located on the distal portion of Xp.
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The gene locus for steroid sulfatase, deficiency of which causes X-linked ichthyosis, is assigned to Xp11 leads to Xpter by analysis of 24 man-Chinese hamster somatic cell hybrids. High steroid sulfatase activity in a hybrid clone having retained only part of Xq is explained by demonstration of an additional late-replicating human X chromosome. This observation confirms previous evidence for noninactivation of the STS locus.
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The segregation of the chromosome 22 markers ACO2, ARSA, and NAGA was studied in somatic cell hybrid clones. These hybrids were isolated following fusion of Chinese hamster (E36 or a3) cells with leucocytes of donors carrying an (X;22) or (1;22) translocation. The results suggest the assignment of ARSA and NAGA to the region 22q13 leads to 22qter and of ACO2 to the region 22q11 leads to 22q13.
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Hybrid cell lines isolated after fusions between Chinese hamster E36 cells and normal human white blood cells were analyzed for human beta-galactosidase isoenzymes and for human chromosomes, especially 3, 12, and 22, the candidates for bearing a beta-galactosidase locus. Results of neuraminidase treatment of the cell lysates and immunological studies showed that in man two structural beta-galactosidase loci are present and can be assigned to chromosomes 3 and 22. No correlation was found between the expression of human beta-galactosidase and the presence of human chromosome 12.
Forty-two primary human-mouse cell hybrids, derived in two separate experiments, were treated with Newcastle disease virus (NDV): eight hybrids were found to produce human interferon and this was shown in every case to be predominantly of the fibroblast type. An extensive analysis was made in terms of karyotype and marker enzymes on all the eight hybrids producing interferon and also on five hybrids which did not produce interferon, five randomly selected hybrids and eleven subclones resistant to diphtheria toxin. The results suggest that, contrary to previous reports, a gene on chromosome 5 is not involved in production of human interferon. Its production was however correlated with the presence of chromosome 9 in the hybrids. Analyses of two sets of human-Chinese hamster hybrid subclones from two different crosses were also consistent with the assignment of a human interferon gene to chromosome 9.
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An immunochemical method has been developed for the quantitative determination of species-specific gene products, for instance alpha-galactosidase and N-acetyl-alpha-galactosaminidase, in man-rodent hybrid cells and in the parental cell lines. Antisera raised against the purified enzymes are covalently coupled to Sepharose 4B. The gene products are specifically removed from a cell lysate by incubating with the appropriate Sepharose-coupled antiserum. After centrifugation followed by washing of the precipitated Sepharose, the enzymic activity can be quantitatively measured on the Sepharose beads. With this technique it has been demonstrated that the ability of human N-acetyl-alpha-galactosaminidase (also known as alpha-galactosidase B) to hydrolyze alpha-galactosidic linkages is lost when the enzyme is expressed in man-Chinese hamster hybrid cells.