摘要
用单功能烷化剂N-甲基-N-硝基-N-亚硝胍(MNNG)作为化学诱变剂处理中国仓鼠卵巢细胞(CHO-K1),使其基因发生点突变,通过含有6-巯基鸟嘌呤(6-TG)的培养基选择,获得次黄嘌呤磷酸核糖转移酶(HPRT,E、C、2、4、2、8)缺陷细胞的克隆.诱发突变频率为1.3×10^(-5).经含次黄嘌呤-氨基喋呤-胸腺嘧啶核苷(HAT)的选择培养基鉴定及大剂量6-TG选择培养基验证,确认选出的克隆为HPRT缺陷型.经6周连续传代,缺陷株表型仍稳定.
Chinese hamster ovary cells (CHO-Kl) were treated with N-methyl-N＇-nitro-N-nitro-soguanidine (MNNG) to cause gene point mutations.The mutagenized cells were subjected to selection in a culture medium containing 6-thioguanine (6-TG).Several cell clones deficient in hypoxanthine-phosphoribosyl-transferase (HPRT) were isolated and cloned.The frequency of mutation at HPRT locus was around 1.3×10^(-5).The cells of the clones picked up died in HAT (hypoxanthine-aminopterin-thymidine) selective medium and survived in a medium containing high concentrations of 6-TG (11μg/ml).These facts further confirmed that these cells were really deficient in HPRT.After continuous culture for six weeks,the HPRT-phenotype was proved to be stable
出处
《军事医学科学院院刊》
CSCD
北大核心
1989年第2期88-91,共4页
Bulletin of the Academy of Military Medical Sciences
关键词
HPRT
缺陷细胞株
诱变
分离
chinese hamster ovary cell
inducing agent
HPRT deficiency cell line
