CELL-CYCLE-SPECIFIC CDNAS FROM MAMMALIAN-CELLS TEMPERATURE SENSITIVE FOR GROWTH
CELL-CYCLE-SPECIFIC CDNAS FROM MAMMALIAN-CELLS TEMPERATURE SENSITIVE FOR GROWTH
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DOI：
10.1073/pnas.81.19.6004
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发表时间：
1984-01-01
期刊：
影响因子：
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通讯作者：
BASERGA, R
中科院分区：
文献类型：
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作者：
HIRSCHHORN, RR;ALLER, P;BASERGA, R
从 ts13 细胞（一种 G1 特异性温度敏感仓鼠细胞系）构建双链 cDNA 文库。克隆到 pBR322 中的 cDNA 是由在 34°C 允许温度下血清刺激后 6 小时从 ts13 细胞分离的聚 (A)+ mRNA 制备的。 C.用从相同细胞制备的G1特异性和G0特异性单链cDNA探针对文库进行差异筛选，鉴定出5个cDNA克隆，其序列优先在G1中表达。 G1 期与这些克隆互补的 RNA 水平比细胞周期其他阶段高 3 至 6 倍。当ts13细胞在39.6℃的限制温度下被阻滞在G1期时。 C，与 p13-2A9 和 p13-4F1 互补的 RNA 水平高达静息群体中的 10 倍，而与 p13-2A8 互补的序列的表达与 G0 中的水平相比没有显着变化。 RNA 和 Southern 凝胶印迹分析表明这些细胞周期特异性克隆代表低拷贝或中等重复的基因序列。细胞周期的另一个 ts 突变体 tsAF8（RNA 聚合酶 II 的 ts 突变体）的结果表明，这些细胞周期特异性序列具有快速更新。使用细胞周期 G1 特异性 ts 突变体提供了一种方法来确定哪些细胞周期依赖性基因与细胞周期进展最相关。
A library of double-stranded cDNA was constructed from ts13 cells, a G1-specific temperature-sensitive hamster cell line. The cDNA, cloned into pBR322, were prepared from poly(A)+ mRNA isolated from ts13 cells 6 h after serum stimulation at the permissive temperature of 34.degree. C. Differential screening of the library with G1-specific and G0-specific single-stranded cDNA probes prepared from the same cells identified 5 cDNA clones whose sequences were preferentially expressed in G1. Levels of RNA complementary to these clones were 3- to 6-fold higher in G1 than in other phases of the cell cycle. When ts13 cells were arrested in G1 at the restrictive temperature of 39.6.degree. C, the levels of RNA complementary to p13-2A9 and p13-4F1 were as high as 10 times that found in a resting population, while the expression of sequences complementary to p13-2A8 did not significnatly change from levels found in G0. RNA and Southern gel blot analysis suggest that these cell-cycle-specific clones represent either low copy or moderately repetitive gene sequences. Results with another ts mutant of the cell cycle, tsAF8, which is a ts mutant of RNA polymerase II, showed that these cell-cycle-specific sequences have a rapid turnover. The use of G1-specific ts mutants of the cell cycle provides an approach to determine which cell-cycle-dependent genes are most relevant to cell-cycle progression.