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Publication : Molecular cloning of transcripts that accumulate during the late G1 phase in cultured mouse cells.

First Author  Nikaido T Year  1991
Journal  Exp Cell Res Volume  192
Issue  1 Pages  102-9
PubMed ID  1984406 Mgi Jnum  J:10882
Mgi Id  MGI:59326 Doi  10.1016/0014-4827(91)90163-o
Citation  Nikaido T, et al. (1991) Molecular cloning of transcripts that accumulate during the late G1 phase in cultured mouse cells. Exp Cell Res 192(1):102-9
abstractText  To identify previously undetected genes that might be involved in later stages of the transition from a quiescent state (G0) to the DNA synthetic phase (S) of murine cells, we set out to isolate cDNA clones derived from mRNAs that appear late in G1 phase in serum-stimulated cells. A lambda-cDNA library was prepared using poly(A)+ RNA from chemically transformed Balb/c 3T3 cells (BP/A31) that had been brought to quiescence and subsequently stimulated for 12 h with serum. From the first screening of approximately 21,000 recombinant phage plaques, about 100 clones were isolated that hybridized to a single-stranded cDNA pool derived from stimulated-cell RNA but not to DNAs made from resting-cell RNA. Eventually, six different clones were identified. The mRNAs from five of these genes increased gradually during the G0 to S transition, in contrast to the immediate-early rise of c-myc mRNA or the later rise of thymidine kinase mRNA. These six clones were sequenced and compared to the GenBank database. Clones LG-80, LG-2, and LG-69 are highly homologous to beta-actin, lactate dehydrogenase, and alpha-tubulin. Clones LG-5, LG-61, and LG-74 had no significant homologies to known sequences. A subtractive cDNA library was used to isolate two additional clones, Sub-S1 and Sub-S2; these have homologies to enolase and ribosomal protein L32. Additional studies that examine the function and regulation of these newly identified late response genes in the pre-DNA synthesis pathway are in progress.
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