鱼类HGPRT缺陷型细胞的诱变和筛选
THE INDUCTON AND SELECTION OF HGPRT-DEFICIENT CELLMUTANTS IN FISH
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摘要: 草鱼卵巢细胞GCG经乙基甲磺酸(EMS)诱变,稳定表达3代以后,采取逐步提高培养基中6-疏基嘌呤(6-MP)的方法,获得对6-MP稳定抗性的细胞,暂定名FMR-1.本文比较了GCG细胞及FMR-1细胞在含6-MP(20μg/ml)和不含6-MP的培养基中生长曲线的特点,并对上述两种细胞的染色体数目和葡萄糖-6-磷酸脱氢酶电泳图谱进行了比较分析.最后对两种细胞在HAT培养基中的生长特点进行了平行对照实验,根据实验结果,可以判定FMR-1细胞为草鱼HGPRT缺陷型细胞.本义还讨论了进一步研究草鱼HGPRT缺陷型细胞的途径及草鱼HGPRT缺陷型细胞在鱼类细胞遗传学上的应用前景.Abstract: The cell line GGG originating from the ovary of grass carp(Ctenpharyngodon idellacur.et val)was treated with ethvl methane sulfonate(EM5)for 72 hours.Aner stable expression of mutation for 3 passages, the eells were treated with 6-mercaptopurine(6-MP)from 4μg/ml to 100μg/ml by stepwise procedures.The isolates did not grow in HAT mediums. Mutants, which have been named FMR-1,and which are deficient in hypoxanthineguanine phosphoribosyl tranferase(HGPRT)were obtained.Identification of biological characteristics of FMR-1 cells showed that chromosomesdecreased in number compared with those of the parental GCG cells. The growth curves of FMR-1 ells and GCG cells in TC-199 mediums which contained 6-MP(20μg/ml)and no 6-MP Were also illustrated and compared. The electrophoretogram of G-6-pD isoenzyme of FMR-1 ells displayed almost the same patterns as that of GCG cells.Further measures to study on the fish HGPRT-deficient cells and possible application ofthese cells are discussed.