【摘要】 本文报道显示鱼类染色体复制带的BrdU-Hoechst-Giemsa方法。采用含20％小牛血清的RPMI-1640培养液进行肾细胞培养,收获前按终浓度加入10μg/ml BrdU和2μg/ml放线菌素D,分别处理16—18小时和1.5—2小时。制片经Hoechst-33258染色,紫外灯照射,2×SSC温育和Gicmsa染色,获得了鲢鱼、鳙鱼和大鳍刺鳑鮍的染色体复制带图象,带纹清晰,结果较稳定。发现BrdU和放线菌素D有抑制鱼类染色体浓缩的作用。更多还原

【Abstract】 A modified BrdU-Hoechst-Giemsa method is described for the demonstration of easily reproducible DNA-replication bands in fish chromosomes. It involves culturing the kidney cells in medium RPMI-1640 supplemented with 20% fetal calf serum at a temperature of 26℃, treating living cells with a final concentration of 10 μg/ml BrdU for 16-18h as well as with 2 μg/ml AMD (actinomycin D) along with 0.05 μg/ml colchi-cine for 1.5-2h before harvest. The air-dried slides are immersed in 1 μg/ml Hoechst 33258 followed by ultraviolet light treatment, incubation in 2XSSC solution and Giem-sa staining. Using this method, the DNA-replication bands in the chromosomes of three species of cyprinid fishes, namely, Hypophthalmichthys molitrix, Aristichthys nobilis, and Acanthorhodeus macropterus, are observed with excellent results. Both BrdU and AMD are found to have effects of inhibiting the condensation of fish chromosomes or chromosomal segments. The problem about the difficulty of revealing Q-, G-, and R-bands in fish chromosomes after applying conventional banding techniques is briefly discussed.更多还原