Hollenberg C P
Chromosoma. 1976 Aug 17;57(2):185-97. doi: 10.1007/BF00292917.
Attention was focussed on the question whether the most active genes of the polytene genome of Chironomus tentans, i.e. rDNA and Balbiani rings, have undergone the same number of duplications as the bulk of the genome. RNA was extracted from isolated salivary glands, enriched for messenger RNA by poly U sepharose chromatography, and labelled with 125Iodine. In situ hybridization showed that, apart from rRNA, Balbiani ring 1 and 2 transcripts are the major hybridizing RNA species present in this enriched preparation. BR1 hybridized only at the b-region. Transcripts of BR6 and puff I 17B are shown to be salivary gland specific and hybridize in situ to an extent of less than 5% of the RNA hybridized to BR1 plus BR2. Saturation hybridization levels of rRNA and Balbiani ring (BR1 plus BR2) RNA with polytene DNA from salivary glands were determined and compared with the saturation levels measured for DNA from adult flies and larvae. rRNA hybridized to the same level (0.09%) with polytene DNA as with DNA from primarily diploid cells (adult fly DNA) or from larvae. An equal hybridization level (0.04%) with each of these three types of DNA was also obtained for BR RNA. The results demonstrate that, during polytenization in C. tentans, the genes for rRNA and BR1 and 2 are duplicated to the same extent as the bulk of the genome.
研究重点关注了摇蚊(Chironomus tentans)多线染色体基因组中最活跃的基因,即核糖体DNA(rDNA)和巴尔比亚尼环,其复制次数是否与基因组的其他部分相同。从分离出的唾液腺中提取RNA,通过聚尿嘧啶琼脂糖层析法富集信使RNA,并用125碘进行标记。原位杂交显示,除了rRNA外,巴尔比亚尼环1和2的转录本是这种富集制剂中主要的杂交RNA种类。BR1仅在b区域杂交。BR6和胀泡I 17B的转录本显示为唾液腺特异性,原位杂交程度不到与BR1加BR2杂交RNA的5%。测定了rRNA和巴尔比亚尼环(BR1加BR2)RNA与唾液腺多线DNA的饱和杂交水平,并与成年果蝇和幼虫DNA的饱和水平进行了比较。rRNA与多线DNA的杂交水平(0.09%)与主要二倍体细胞(成年果蝇DNA)或幼虫DNA的杂交水平相同。BR RNA与这三种类型DNA中的每一种的杂交水平也相同(0.04%)。结果表明,在摇蚊多线化过程中,rRNA以及BR1和2的基因与基因组的其他部分以相同程度进行了复制。