Genetic analysis with man-mouse somatic cell hybrids. Linkage between human lactate dehydrogenase A and B and peptidase B.
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Biomedical subjects
Publications and source records attributed to R J Klebe.
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The ES-2 esterase is a kidney-associated esterase and is expressed in a cloned mouse renal adenocarcinoma cell line. Somatic cell hybridization of the mouse renal adenocarcinoma with either mouse or human fibroblasts leads to the marked reduction of ES-2 esterase titer. The extinction of ES-2 esterase activity in mousexhuman hybrids is reversible. Extinction correlates with the presence of the human C(10) chromosome in the somatic cell hybrids; whereas reexpression of the ES-2 esterase is observed in hybrids which have lost C(10). Thus, the proposed regulator element involved is probably structurally linked to C(10); however, linkage to a member of the D or G groups cannot be completely excluded.
The techniques described permit the controlled production of large numbers of proliferating somatic cell hybrids in a relatively short period of time. Sendai virus is used to promote cell hybridization. beta-propriolactone is employed as the inactivating agent of Sendai virus since it produces complete loss of viral infectivity while preserving viral fusion capacity. Cells are fused in monolayer, instead of in suspension, since fixing cells in two dimensions permits one to control cell contacts during the fusion event through the expedient of varying multiplicities of the parental cells and the total cell density. Under the conditions described, a several hundred fold increase in the number of hybrid clones obtained is seen as compared to the controls.