Elsevier S M, Ruddle F H
Chromosoma. 1975 Oct 14;52(3):219-28. doi: 10.1007/BF00332112.
Cytological detection of cistrons coding for 18S and 28S ribosomal RNA (rRNA) within the genome of Mus musculus inbred strain SEC/1ReJ was accomplished using the technique of in situ hybridization. Metaphase chromosome spreads prepared from cultured fetal mouse cells were stained with quinacrine-HCl and photographed. After destaining, they were hybridized to Xenopus laevis tritiated 18S and 28S rRNA, specific activity 7.5 X 10(6) dpm/mug. Silver grains clustered over specific chromosomes were readily apparent after 4 months of autoradiographic exposure. The identity of the labelled chromosomes was established by comparing the autoradiographs to quinacrine photographs showing characteristic fluorescent banding of the chromosomes in each metaphase spread. The 18S and 28S rRNA was found to hybridize to chromosomes 12, 18, and 16. Statistical analysis of the grain distribution over 26 spreads revealed that the three chromosomes were significantly labelled. Grains over these chromosomes were concentrated in an area immediately distal to the centromere, a region which in chromosomes 12 and 18 in this particular strain is the site of a secondary constriction. The relative size of the secondary constrictions, long and thus prominent on chromosome 12, obvious but shorter on 18, and indistinguishable on chromosome 16, correlated with the average number of grains observed over the centromeric region of these chromosomes, 2.5, 1.0, and 0.78, respectively.
利用原位杂交技术完成了对小家鼠近交系SEC/1ReJ基因组中编码18S和28S核糖体RNA(rRNA)的顺反子的细胞学检测。用培养的胎鼠细胞制备中期染色体铺片,用盐酸喹吖因染色并拍照。脱色后,将其与特异活性为7.5×10⁶ dpm/μg的非洲爪蟾氚化18S和28S rRNA杂交。经过4个月的放射自显影曝光后,聚集在特定染色体上的银颗粒清晰可见。通过将放射自显影片与显示每个中期铺片中染色体特征荧光带型的喹吖因照片进行比较,确定了标记染色体的身份。发现18S和28S rRNA与12号、18号和16号染色体杂交。对26个铺片上颗粒分布的统计分析表明,这三条染色体有明显标记。这些染色体上的颗粒集中在着丝粒紧邻的区域,在这个特定品系中,12号和18号染色体上的这个区域是次缢痕的位置。次缢痕的相对大小,在12号染色体上长且明显,在18号染色体上明显但较短,在16号染色体上难以区分,与在这些染色体着丝粒区域观察到的颗粒平均数相关,分别为2.5、1.0和0.78。