PUBLICATION
Preparation of biologically active monomeric recombinant zebrafish (Danio rerio) and rainbow trout (Oncorhynchus mykiss) recombinant growth hormones
- Authors
- Oc?o?, E., Solomon, G., Hayouka, Z., Gertler, A.
- ID
- ZDB-PUB-180520-4
- Date
- 2018
- Source
- Fish physiology and biochemistry 44(4): 1215-1222 (Journal)
- Registered Authors
- Keywords
- Fish, Growth hormone, Rainbow trout, Recombinant, Zebra fish
- MeSH Terms
-
- Rabbits
- Oncorhynchus mykiss/metabolism*
- Growth Hormone/genetics
- Growth Hormone/isolation & purification
- Growth Hormone/metabolism*
- Receptors, Somatotropin/genetics
- Receptors, Somatotropin/metabolism*
- Zebrafish/metabolism*
- Humans
- Animals
- Sheep
- PubMed
- 29777415 Full text @ Fish Physiol. Biochem.
Citation
Oc?o?, E., Solomon, G., Hayouka, Z., Gertler, A. (2018) Preparation of biologically active monomeric recombinant zebrafish (Danio rerio) and rainbow trout (Oncorhynchus mykiss) recombinant growth hormones. Fish physiology and biochemistry. 44(4):1215-1222.
Abstract
Fish growth hormones (GHs) play an important role in regulating growth, metabolism, reproduction, osmoregulation, and immunity and have thus garnered attention for their application in aquaculture. Zebrafish GH (zGH) cDNA or rainbow trout GH (rtGH) cDNA was cloned into the pMon3401 vector, expressed in MON105-competent Escherichia coli and purified to homogeneity. Their biological activity was evidenced by their ability to interact with ovine GH receptor extracellular domain and stimulate GH receptor-mediated proliferation in FDC-P1-3B9 cells stably transfected with rabbit GH receptor. The relative affinity of zGH and rtGH, estimated by IC50, was about 38-fold and 512-fold lower, respectively, than ovine GH. This is likely the reason for the low biological activity in cells with rabbit GH receptor, ~ 36-fold lower for zGH and ~ 107-fold lower for rtGH than for human GH. This was not due to improper refolding, as evidenced by circular dichroism analysis. Predicting the activity of fish GHs is problematic as there is no one single optimal in vitro bioassay; heterologous assays may be ambiguous, and only homologous assays are suitable for measuring activity.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping