犬肾传代细胞HGPRT缺失株的诱变  

Induced Mutation of HGPRT-deficient Mading-Darby Ca-nine Kidney Cell(MDCK)

作　　者：钟志宏 刘兴发 娄淑杰 杨春梅 夏咸柱[1] 刘家友 

机构地区：[1]兰州军区军事医学研究所,解放军农牧大学军事兽医研究所

出　　处：《中国兽医学报》1995年第3期255-259,共5页Chinese Journal of Veterinary Science

基　　金：军队青年基金

摘　　要：应用８－氮鸟嘌呤（８－Ａｚａｇｕｉｚｉｎｅ），通过大剂量追加细胞法、剂量递增法和紫外线预先照射交替培养法，共获得９株犬肾传代细胞系（ＭＤＣＫ）抗药细胞（剂量分别为２０ｕｇ／ｍＬ和５０ｕｇ／ｍＬ），经ＨＡＴ系统敏感性试验和６－疏鸟嘌呤抗性鉴定，其中有８株为犬肾传代细胞ＨＧ－ＰＲＴ缺失突变株（ＭＤＣＫ／ＨＧＰＲＴ－）。该突变株细胞Ｇｉｅｍｓａ染色后呈卵圆形，核呈圆形，８３世代细胞（ＭＤＣＫ８３／ＨＧＰＲＴ－）在分种后２４～９６ｈ为细胞分裂的对数生长期，其群体倍增时间为１８．７ｈ，细胞染色体众数为７９条，呈现近二倍体核型，该细胞对ＩＣＨＶ的ＴＣＩＤ５０为１０４．９／ｍＬ。这些指标与诱变前细胞相似。Nine clones of 8-azaguanine-resistant MDCK were obtained by three in-duced mutation methods in this experiment.All these cells were able to grow in thepresence of a second purine analogy,6-thioguanine but only eight of them were not ableto grow in counter-selective medium containing hypoxanthine,aminopterin and thymi-dine(HAT).The eight clones of drug-resistant MDCK were identified as HGPRT-defi-cient MDCK,desingnated MDCK/NCPRT-.The MDCK/HGPRT-maintained an ep-ithelial-like appearance. The growth curve of these cells at 83rd passage showed a loga-rithmic increase in the number of cells between 24 and 96 hours and population doublingtime was estimated to be 18. 7 hours. Karyotypie analysis showed the total in chromo-some number was 79.The MDCK/HGPRT- was still susceptible to infection of infec-tous canine hepatitis virus and TCID50 was 104.9/mL.

关 键 词：犬 犬肾传代细胞 HGPRT 缺失株 诱变 

分 类 号：S829.2[农业科学—畜牧学]