Preparation of a subtractive cDNA library enriched in cDNAs which expressed at a high level in cultured senescent human fibroblasts.
Subtracted cDNA library was prepared by subtracting [cDNA from young growing SV40-transformed human fibroblasts] from [cDNA from growing SV40-transformed fibroblasts in extended lifespan]. Isolated cDNA clones which expressed at high level in life-extended transformed cells also expressed at high level in normal senescent fibroblasts but did at low level in growing and growth-arrested young cells. Neither fibronectin nor procollagen cDNA was isolated. This cDNA library is useful for isolation of senescent-specific cDNA species which express at high level in normal senescent cells but at low level in growing and growth-arrested young cells, avoiding growth-arrest-specific cDNAs.
Tahara
H
H
Department of Cellular and Molecular Biology, Hiroshima University School of Medicine, Japan.
Hara
E
E
Tsuyama
N
N
Oda
K
K
Ide
T
T
D000818:Animals; D015152:Blotting, Northern; D002417:Cattle; D002455:Cell Division; D002461:Cell Line, Transformed; D016922:Cellular Senescence; D002999:Clone Cells; D018076:DNA, Complementary / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D005333:Fetus; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D005353:Fibronectins / Q000096:biosynthesis; D015870:Gene Expression; D015723:Gene Library; D006801:Humans; D011347:Procollagen / Q000096:biosynthesis; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism; D013539:Simian virus 40 / Q000235:genetics
D018076:DNA, Complementary; D005353:Fibronectins; D011347:Procollagen; D012313:RNA