Isolation and expression of tilapia (Oreochromis niloticus) serine 8-type GnRH coding and regulatory sequences.
The complete Serine 8-type gonadotropin releasing hormone (GnRH) coding sequence with a substantial 5-prime regulatory sequence (5 kb) has been isolated and characterised in Nile Tilapia (Oreochromis niloticus) from a relevant genomic library. The primary structure of the protein precursor was identified for this gene. The promoter efficacy has been tested using 0.6 kb of the GnRH promoter driving a lacZ reporter gene in both cultured spleen cells and transiently expressing zebrafish. In the cell transfection experiments, the average level of beta-galactosidase activity in transfected cells was more than 2.1 (P<0.05) times higher than the control promoter-less vector in five independent cultures indicating that the 0.6 GnRH/lacZ construct is able to express in spleen cells. In addition, the transient expression of the lacZ gene was detected in the brain of G0 zebrafish embryos (Danio rerio) 4 days after fertilisation following egg injection with the construct, which demonstrated the efficacy of the tilapia GnRH promoter.
Farahmand
Hamid
H
University of Southampton, School of Biological Sciences, Division of Cell Sciences, Biomedical Sciences Building, Bassett Crescent East, Southampton SO16 7PX, UK.
Rahman
M Azizur
MA
Sohm
Frédéric
F
Hwang
Gyu-Lin
GL
Maclean
Norman
N
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004247:DNA / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D004625:Embryo, Nonmammalian / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D007987:Gonadotropin-Releasing Hormone / Q000235:genetics*; D008297:Male; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D012689:Sequence Homology, Nucleic Acid; D017210:Tilapia / Q000235:genetics*; D014162:Transfection; D015027:Zebrafish; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D011993:Recombinant Fusion Proteins; D007987:Gonadotropin-Releasing Hormone; D004247:DNA; D001616:beta-Galactosidase