Building comparative gene expression databases for the mouse preimplantation embryo using a pipeline approach to UniGene.
To understand early mammalian development there is a need to compare profiles of gene expression from different stages of the preimplantation mouse embryo. We describe here a method that uses gene expression data held in the UniGene database of the National Institutes of Health (NIH). The full mouse UniGene database (build #151) contains 43,104 gene clusters generated from approximately 4.1 million sequences. The Expressed Sequence Tags (EST) used to build UniGene are derived from cDNA libraries that are archived separately in the database of Expressed Sequence Tags (dbEST) database, with their own catalogue numbers. The mouse dbEST database contains 32 non-normalized dbEST libraries constructed from preimplantation stages (unfertilized oocyte, fertilized oocyte, 2-, 4-, 8- and 16-cell embryo and blastocyst). These libraries contain 219,852 EST sequences mapping to 15,731 UniGene clusters. We have developed a computational pipeline approach that imports and aggregates inventories of gene expression contained in these dbEST libraries. It uses these data to build an annotated web-based database of preimplantation gene expression with an in-built capacity for comparison of expression profiles. Comparison of gene expression profiles obtained for each developmental stage show statistically significant changes in gene expression during preimplantation development. These in silico-generated profiles were validated using RT-PCR.
Stanton
J-A L
JA
Anatomy and Structural Biology, University of Otago, Dunedin, New Zealand. jo.stanton@anatomy.otago.ac.nz
Macgregor
A B
AB
Mason
C
C
Dameh
M
M
Green
D P L
DP
D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D019295:Computational Biology / Q000379:methods*; D030541:Databases, Genetic; D020224:Expressed Sequence Tags; D020869:Gene Expression Profiling / Q000379:methods*; D051379:Mice; D020133:Reverse Transcriptase Polymerase Chain Reaction