IDENTIFICATION OF A SET OF GENES EXPRESSED DURING THE G0/G1 TRANSITION OF CULTURED MOUSE CELLS
IDENTIFICATION OF A SET OF GENES EXPRESSED DURING THE G0/G1 TRANSITION OF CULTURED MOUSE CELLS
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DOI：
10.1002/j.1460-2075.1985.tb04057.x
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发表时间：
1985-01-01
期刊：
影响因子：
11.4
通讯作者：
NATHANS, D
中科院分区：
文献类型：
--
作者：
LAU, LF;NATHANS, D
为了确定以前未检测到的基因，可能参与从静止状态（G 0）的哺乳动物细胞的增殖状态（G1）的过渡，我们着手分离来自mRNA的cDNA克隆出现在血清刺激的细胞中的蛋白质合成的情况下。A.利用BALB/c 3 T3细胞的poly（A）+ RNA制备cDNA文库，该细胞已被静止并随后在放线菌酮存在下用血清刺激。筛选了约50000个重组噬菌体噬斑，分离出357个克隆，它们与来自刺激细胞RNA的探针杂交，但不与来自静息细胞RNA的探针杂交。交叉杂交分析表明，四个RNA序列家族占约。90%的克隆人其中一个克隆与肌动蛋白探针杂交;没有一个与测试的13种癌基因探针杂交。五种不同的RNA，似乎是以前未表征的已被进一步分析。这些RNA在血清或纯化的生长因子或肿瘤促进剂刺激后迅速积累和衰减，并且它们在放线菌酮存在下被血清超诱导。三种RNA可以通过与cDNA杂交而富集，并在体外翻译，产生λ蛋白。43、40和35 kd。
To identify previously undetected genes that may be involved in the transition from a resting state (G0) to a proliferative state (G1) of mammalian cells, we set out to isolate cDNA clones derived from mRNAs that appear in serum-stimulated cells in the absence of protein synthesis. A .lambda. cDNA library was prepared using poly(A)+ RNA from BALB/c 3T3 cells that had been brought to quiescence and subsequently stimulated with serum in the presence of cycloheximide. Approximately 50000 recombinant phage plaques were screened, and 357 clones were isolated that hybridized to probes derived from stimulated-cell RNA but not to probes from resting-cell RNA. Cross hybridization analysis showed that four RNA sequence families account for .apprx. 90% of these clones. One of the clones hybridized to an actin probe; none hybridized to any of 13 oncogene probes tested. Five different RNAs that appear to be previously uncharacterized have been further analyzed. These RNAs accumulate and decay rapidly following stimulation by serum or purified growth factors, or by a tumor promoter, and they are superinduced by serum in the presence of cycloheximide. Three of the RNAs could be enriched by hybridization to cDNAs and translated in vitro, yielding proteins of .apprx. 43, 40 and 35 kd, respectively.