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In situ hybridization screen in zebrafish for the selection of genes encoding secreted proteins
Author(s) -
Crosier Philip S.,
Bardsley Anne,
Horsfield Julia A.,
Krassowska Anna K.,
Lavallie Edward R.,
CollinsRacie Lisa A.,
Postlethwait John H.,
Yan YiLin,
Mccoy John M.,
Crosier Kathryn E.
Publication year - 2001
Publication title -
developmental dynamics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.634
H-Index - 141
eISSN - 1097-0177
pISSN - 1058-8388
DOI - 10.1002/dvdy.1218
Subject(s) - biology , zebrafish , encode , gene , in situ hybridization , genetics , expressed sequence tag , complementary dna , computational biology , gene expression
An in situ hybridization expression screen using a signal sequence trap system has been conducted in zebrafish to isolate cDNAs that encode secreted proteins. Random clones (secreted expressed sequence tags; sESTs) were sequenced from zebrafish embryonic (18–24 hr postfertilization) and adult kidney libraries. From the two RNA sources, 627 random sEST cDNAs were identified as being homologous or identical to known genes and 166 clones encode currently unidentified genes. The sESTs represent a broad range of enzymes and other regulatory molecules. Whole‐mount in situ hybridization analysis was carried out by using antisense probes generated from 244 selected sESTs, and a range of expression patterns was obtained. Genetic mapping undertaken with sEST sequences demonstrated that assignment of map position was attainable by using 5′ primers. The signal sequence trap system used in this work has yielded a range of cDNAs that encode secreted proteins and, together with analysis of patterns of expression and genetic mapping, has the potential to facilitate analysis of signaling pathways central to development and physiology. © 2001 Wiley‐Liss, Inc.