Steinemann M
Chromosoma. 1981;82(2):267-88. doi: 10.1007/BF00286111.
DNA fiber autoradiography was used to determine parameters underlying the DNA replication of the eukaryotic chromosome in Drosophila diploid brain cells organ culture. The average rate of fork movement, estimated from 4 different labelling intervals, is 0.35 micron/min at 25 degrees C. Of the tandem arrays 93% show patterns which are compatible with bidirectional replication, 7% show unidirectional replication. The unidirectional mode of replication is interpreted as being a consequence of the experimental schedule (using hot-cold pulse labelling) combined with the occurrence of termination signals. - Some autoradiograms showed the expected two grain tracks of different densities; others showed only a high density track. The latter were most prominent in arrays of short replicons (less than 10 micron) which correlate with replicating satellite sequences. - The majority of replicons fall into size classes less than 100 micron. The frequency distribution is skewed towards larger replicon sizes; it spans 2-238 micron, has a mean of x = 25.6 micron and a median of approximately 21.0 micron. If the distribution is corrected for supposed satellite replicons, the median increases of approximately 31.0 micron. - In experiments using warm-hot pulse labelling, arrays were scored which must have been a consequence of fixed termination signals. Furthermore, grain tracks diverging from weak labelled centers often have different lengths, indicating that these replicons contain two diverging replicating sections of unequal length.
利用DNA纤维放射自显影技术来确定果蝇二倍体脑细胞器官培养中真核染色体DNA复制的潜在参数。在25摄氏度下,根据4个不同的标记间隔估算出的叉形移动平均速率为0.35微米/分钟。在串联阵列中,93%呈现出与双向复制相符的模式,7%呈现单向复制。单向复制模式被解释为实验方案(使用热-冷脉冲标记)与终止信号出现相结合的结果。- 一些放射自显影片显示出预期的两条不同密度的颗粒轨迹;另一些则只显示出高密度轨迹。后者在与卫星序列复制相关的短复制子(小于10微米)阵列中最为突出。- 大多数复制子的大小类别小于100微米。频率分布向较大的复制子大小倾斜;其范围为2-238微米,平均值为x = 25.6微米,中位数约为21.0微米。如果对假定的卫星复制子的分布进行校正,中位数增加到约31.0微米。- 在使用温-热脉冲标记的实验中,对一些阵列进行了评分,这些阵列肯定是固定终止信号的结果。此外,从弱标记中心发散的颗粒轨迹往往长度不同,这表明这些复制子包含两个长度不等的发散复制部分。