title: Use of the SST-REX method for the identification of genes expressed at the condensation stage of chondrogenic cell line ATDC5.
abstract: In endochondral ossification, chondrogenesis precedes bone development. Cartilage differentiation is initiated by the formation of chondrogenic cell condensates. Thus, it is essential to investigate genes expressed at the condensation stage for a better understanding of chondrogenesis and ossification. To this end, we constructed a cDNA library from the mRNA fraction derived from a chondrogenic cell line (ATDC5) at the cell condensation stage by using the signal sequence trap by retrovirus-mediated expression (SST-REX) method. We obtained 486 factor (IL-3)-independent clones by screening 5.7 x 10(3) clones. DNA sequencing analysis of the clones identified genes encoding 157 known proteins and 4 novel proteins. These 4 genes encoding novel proteins were expressed not only in chondrogenic ATDC5 cells but also in osteogenic MC3T3-E1 and myogenic C2C12 cells. The mRNA expression level of 1 of the 4 clones increased at the calcification stage of the differentiation of MC3T3-E1 cells. The results demonstrate that the SST-REX method is a useful experimental system to identify genes involved in the complicated mechanisms of bone formation.
authors.affiliation: Research Institute for Biological Sciences, Tokyo University of Science (Rikadai), 2669 Yamazaki, Noda, 278-0022, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Noguchi
authors.forename: Naoko
authors.initials: N
authors.lastname: Watanabe
authors.forename: Ryoji
authors.initials: R
authors.lastname: Fujimaki
authors.forename: Toshio
authors.initials: T
authors.lastname: Kitamura
authors.forename: Yoshihide
authors.initials: Y
authors.lastname: Hayashizaki
authors.forename: Shigeru
authors.initials: S
authors.lastname: Miyaki
authors.forename: Kenichi
authors.initials: K
authors.lastname: Tezuka
authors.forename: Nobumichi
authors.initials: N
authors.lastname: Hozumi
mesh_terms: D000818:Animals; D015152:Blotting, Northern; D001842:Bone and Bones / Q000378:metabolism; D002356:Cartilage / Q000378:metabolism*; D002454:Cell Differentiation; D002460:Cell Line; D019902:Chondrocytes / Q000378:metabolism; D020219:Chondrogenesis; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D015870:Gene Expression; D015723:Gene Library; D007377:Interleukin-3 / Q000378:metabolism; D051379:Mice; D012313:RNA / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D012190:Retroviridae / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D007377:Interleukin-3; D012333:RNA, Messenger; D012313:RNA