Isolation of developmentally regulated novel genes based on sequence identity and gene expression pattern.
Based on the surmise that a variety of genes might play important roles in embryonic development and tissue differentiation, and that some of them are likely to be expressed in undifferentiated ES cells, we attempted to identify new genes from the ES cell cDNA library. The modified method of expressed sequence tags (ESTs) and the examination of the expression patterns in adult tissues and in vitro differentiated ES cells were utilized in this study. We have isolated and identified several novel cDNA clones with interesting developmental expression pattern. Among the 83 clones randomly chosen, 23 clones (27.7%) have no homology to any sequences in public databases. The rest contain limited or complete sequence homology to the previously reported mammalian genes or ESTs, yet some clones have not been previously identified in the mouse. To examine the expression profile of clones during development and differentiation, sets of slot blots were hybridized with developmental stage specific or tissue specific probes. Out of 40 novel clones tested (21 totally unknown clones and 19 unidentified clones in mouse), most of them were up- or down-regulated as differentiation proceeded, and some clones showed differentiation-stage specific expression profiles. Surprisingly, a majority of genes were also expressed in adult tissues, and some clones even revealed tissue specific expression. These results demonstrate that not only was the strategy we employed in this study quite efficient for screening novel genes, but that the information gained by such studies would also be a useful guide for further analysis of these genes. It also suggests the feasibility of this approach to explore the genomewide network of gene expression during complicated biological processes, such as embryonic development and tissue differentiation.
Kim
S J
SJ
Department of Life Science, College of Natural Sciences, Hanyang University, Seoul, Korea.
Shin
J H
JH
Kim
J
J
Kim
S H
SH
Chae
J H
JH
Park
E J
EJ
Seong
R H
RH
Hong
S H
SH
Park
S D
SD
Jeong
S
S
Kim
C G
CG
D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002454:Cell Differentiation; D002460:Cell Line; D006224:Cricetinae; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D016208:Databases, Factual; D020224:Expressed Sequence Tags; D018507:Gene Expression Regulation, Developmental; D005796:Genes / Q000235:genetics*; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D051381:Rats; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary