Corsaro C M, Migeon B R
Somatic Cell Genet. 1978 Sep;4(5):531-40. doi: 10.1007/BF01542924.
Single-step mutations to ouabain resistance have been induced in Lesch-Nyhan diploid fibroblasts carrying a variant G6PD isozyme (OuaR/HGPRT-/G6PDA). Two of these mutant strains, differing in the degree of ouabain resistance, were fused with wild-type human diploid fibroblasts (OuaS/HGPRT+/G6PDB) and hybrids isolated in doubly selective medium, ouabain + HAT (HOT). Four hybrids, identified by G6PD heteropolymers and stable tetraploid karyotypes, expressed ouabain resistance intermediate to that of the parents. The degree of ouabain resistance was influenced by the specific mutant allele of the OuaR parent, indicating both mutant and wild-type alleles are simultaneously expressed in hybrid cells. Intraspecific hybrids of this kind are especially suited to studies of gene regulation because all relevant genes present and the number of alleles at each locus is known.
在携带变异型葡萄糖-6-磷酸脱氢酶同工酶(OuaR/HGPRT-/G6PDA)的莱施-奈恩二倍体成纤维细胞中,已诱导出对哇巴因抗性的单步突变。其中两个在哇巴因抗性程度上不同的突变株,与野生型人二倍体成纤维细胞(OuaS/HGPRT+/G6PDB)融合,并在双选择培养基(哇巴因+HAT,即HOT)中分离出杂种细胞。通过葡萄糖-6-磷酸脱氢酶异聚物和稳定的四倍体核型鉴定出的四个杂种细胞,其表达的哇巴因抗性介于亲本之间。哇巴因抗性程度受OuaR亲本的特定突变等位基因影响,表明突变型和野生型等位基因在杂种细胞中同时表达。这种种内杂种特别适合用于基因调控研究,因为所有相关基因都存在,且每个基因座的等位基因数量是已知的。