Sederoff R, Lowenstein L
Cell. 1975 Jun;5(2):183-94. doi: 10.1016/0092-8674(75)90026-4.
Long pyrimidine tracts, purified from Drosophila melanogaster DNA after treatment with formic acid-diphenylamine, were used as template for E. coli RNA polymerase to produce a polynucleotide containing only purines. This polypurine RNA hybridized specifically to D. melanogaster DNA with high efficiency at low Cot values. The resulting hybrid showed high thermal stability. When polypurine RNA was subjected to complete hydrolysis with ribonuclease T1, over 90% of the nucleotide products were ApGp and ApApGp. Partial hydrolysis yielded a distinct additional component, ApApGpApGp + ApGpApApGp. We conclude that the major sequence in the polypurine transcript is (ApGpApApGp)n. In situ hybridization to salivary gland polytene chromosomes and to metaphase chromosomes from neural ganglia indicated that polypyrimidines complementary to polypurine RNA are located in heterochromatin. In femal cells, the predominant labeling was on centromeric heterochromatin of the 2nd chromosome. We have verified the location of polypyrimidines in neural ganglion cells, by using a cytological marker of chromosomes 2. In male cells, hybrid was also found on the Y chromosome.
从经甲酸 - 二苯胺处理后的黑腹果蝇DNA中纯化得到的长嘧啶序列,被用作大肠杆菌RNA聚合酶的模板,以产生仅含嘌呤的多核苷酸。这种多嘌呤RNA在低Cot值下能高效地与黑腹果蝇DNA特异性杂交。所得杂交体显示出高热稳定性。当多嘌呤RNA用核糖核酸酶T1进行完全水解时,超过90%的核苷酸产物是ApGp和ApApGp。部分水解产生了一种独特的额外成分,即ApApGpApGp + ApGpApApGp。我们得出结论,多嘌呤转录本中的主要序列是(ApGpApApGp)n。对唾液腺多线染色体和神经节中期染色体进行原位杂交表明,与多嘌呤RNA互补的多嘧啶位于异染色质中。在雌性细胞中,主要标记位于第二条染色体的着丝粒异染色质上。我们通过使用第二条染色体的细胞学标记,验证了神经节细胞中多嘧啶的位置。在雄性细胞中,在Y染色体上也发现了杂交体。