H.S. Wang

Localization of the gene encoding human Factor V to chromosome 1q21–25

The gene encoding human coagulation Factor V (FV), one of the cofactors in the blood clotting process, has been mapped to chromosome 1 by both Southern hybridization to DNA from human-hamster somatic cell hybrids and in situ hybridization. The whole plasmid pUC3A containing a 1.5-kb cDNA sequence for FV was 32P-labeled for Southern analysis and 3H-labeled for in situ hybridization to metaphase chromosomes. The results localized the FV gene to the region of 1q21-25.

Regional localization of 18 human X-linked DNA sequences

A series of human probes with unique sequences has been isolated from a recombinant phage library constructed with DNA obtained from a human-hamster hybrid cell line. This cell line contained the X chromosome as the only human component. For 18 of these probes, a human X-chromosome origin has been confirmed and they have been regionally assigned by a combination of techniques: dosage studies utilizing DNA from human fibroblasts carrying X-chromosome duplications and deletions; the presence or absence of hybridization to digested DNA from hybrid lines carrying fragments of the X chromosome; and in situ hybridization to metaphase chromosomes. The use of dosage as a means to regionally assign probes significantly improves resolution of the X chromosome.

Assignment of the sorbitol dehydrogenase locus to human chromosome 15 pter→q21

The locus for sorbitol dehydrogenase (SORD, E.C. 1.1.1.14) has been shown to segregate with hexosaminidase A and mannose phosphate isomerase in a series of human-Chinese hamster somatic cell hybrids. Cytogenetic analysis supports the assignment to chromosome 15 and further defines the gene locus to the region 15pter→q21.

Cytogenetic characteristics of 26 polyethylene glycol-induced human-hamster hybrid cell lines.

A cytological analysis of 26 polyethylene glycol (PEG) induced human/hamster hybrid lines has shown that such lines are similar to inactivated Sendai virus (ISV) induced hybrids in respect to stability, retention of specific chromosomes, and cell selection. The evolution of stable hybrid cell lines carrying variable human chromosome complements depends upon a balance being established between the retained human and hamster genomes. This balance is a result of random loss of human and hamster chromosomes followed by selection of the fittest stem lines. A major mechanism ofchromosome loss may be fragmentation and elimination of acentric fragments. Twelve of the 26 lines had stabilized by the 30th passage, an incidence similar to that found with ISV-induced hybrids studied in this laboratory. Thus, PEG may be considered to be an ideal chemical for inducing somatic cell hybrids for genetic analysis.

The effect of gamma irradiation of human cells and somatic cell hybridization on sister chromatid exchange frequencies in the hamster chromosomes of hamster-human hybrid lines

Sister chromatid exchange (SCE) frequency was studied in human/hamster hybrid lines in which the human cells had been exposed to gamma irradiation at doses of 0, 10, 20, and 40 grays. A higher SCE frequency was found in the hamster chromosomes in the hybrid lines than was found in the hamster parental cell line CHW1103. There was also a dose-dependent increase of SCE frequency in the unirradiated hamster chromosomes in the hybrid lines in which the human parental cells had been irradiated.

Human chromosomes control growth of human-chinese hamster somatic cell hybrids

Several independent human-Chinese hamster hybrid cell lines have been monitored for growth rate and chromosome composition. The rate of growth, as indicated by doubling time during log phase, was positively correlated with the number of human chromosomes, and human chromosomes 3 and 21 were present in stable hybrid cell lines more frequently than expected.