A high throughput screen to identify secreted and transmembrane proteins involved in Drosophila embryogenesis.
Secreted and transmembrane proteins play an essential role in intercellular communication during the development of multicellular organisms. Because only a small number of these genes have been characterized, we developed a screen for genes encoding extracellular proteins that are differentially expressed during Drosophila embryogenesis. Our approach utilizes a new method for screening large numbers of cDNAs by whole-embryo in situ hybridization. The cDNA library for the screen was prepared from rough endoplasmic reticulum-bound mRNA and is therefore enriched in clones encoding membrane and secreted proteins. To increase the prevalence of rare cDNAs in the library, the library was normalized using a method based on cDNA hybridization to genomic DNA-coated beads. In total, 2,518 individual cDNAs from the normalized library were screened by in situ hybridization, and 917 of these cDNAs represent genes differentially expressed during embryonic development. Sequence analysis of 1,001 cDNAs indicated that 811 represent genes not previously described in Drosophila. Expression pattern photographs and partial DNA sequences have been assembled in a database publicly available at the Berkeley Drosophila Genome Project website (). The identification of a large number of genes encoding proteins involved in cell-cell contact and signaling will advance our knowledge of the mechanisms by which multicellular organisms and their specialized organs develop.
Kopczynski
C C
CC
Department of Molecular and Cell Biology, Howard Hughes Medical Institute, University of California, Berkeley, CA 94720-3200, USA.
Noordermeer
J N
JN
Serano
T L
TL
Chen
W Y
WY
Pendleton
J D
JD
Lewis
S
S
Goodman
C S
CS
Rubin
G M
GM
D000818:Animals; D003195:Computer Communication Networks; D018076:DNA, Complementary / Q000235:genetics; D016208:Databases, Factual; D004330:Drosophila / Q000196:embryology* / Q000235:genetics* / Q000502:physiology; D018507:Gene Expression Regulation, Developmental; D017344:Genes, Insect; D017403:In Situ Hybridization; D019476:Insect Proteins / Q000235:genetics* / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000302:isolation & purification
D018076:DNA, Complementary; D019476:Insect Proteins; D008565:Membrane Proteins; D012333:RNA, Messenger