Toward the gene catalogue of sea urchin development: the construction and analysis of an unfertilized egg cDNA library highly normalized by oligonucleotide fingerprinting.
We describe the use of oligonucleotide fingerprinting for the generation of a normalized cDNA library from unfertilized sea urchin eggs and report the preliminary analysis of this library, which resulted in the establishment of a partial gene catalogue of the sea urchin egg. In an analysis of 21,925 cDNA clones by hybridization with 217 oligonucleotide probes, we were able to identify 6291 clusters corresponding to different transcripts, ranging in size from 1 to 265 clones. This corresponds to an average 3.5-fold normalization of the starting library. The normalized library represents about one-third of all genes expressed in the sea urchin egg. To generate sequence information for the transcripts represented by the clusters, representative clones selected from 711 clusters were sequenced. The construction and preliminary analysis of the normalized library are the first steps in the assembly of an increasingly complete collection of maternal genes expressed in the sea urchin egg, which will provide a number of insights into the early development of this well-characterized model organism.
Max-Planck-Institut für Molekulare Genetik, Ihnestrasse 73, Berlin, 14195, Germany. poustka@mpimg-berlin-dahlem.mpg.de
A J
AJ
Poustka
R
R
Herwig
A
A
Krause
S
S
Hennig
S
S
Meier-Ewert
H
H
Lehrach
D000818:Animals; D003001:Cloning, Molecular; D016172:DNA Fingerprinting; D018076:DNA, Complementary / Q000737:chemistry / Q000145:classification / Q000235:genetics; D018507:Gene Expression Regulation, Developmental; D015723:Gene Library; D008969:Molecular Sequence Data; D009841:Oligonucleotides; D010063:Ovum / Q000737:chemistry / Q000254:growth & development / Q000378:metabolism*; D012617:Sea Urchins / Q000737:chemistry / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
D018076:DNA, Complementary; D009841:Oligonucleotides