Recombinant Carp (Cyprinus carpio) Growth Hormone: Expression, Purification, and Determination of Biological Activity in Vitro and in Vivo

Cytokines in fish: an update

“…With the clear structural similarities to cytokines that many hormones possess, it should not be forgotten that several fish hormones are sequenced and recombinant proteins exist, as with growth hormone (Fine et al, 1993;MartinezBarbera et al, 1994) which has already been shown to modulate immune responses in fish such as phagocyte respiratory burst activity (Sakai et al, 1995) and allograft rejection (Nevid & Meier, 1995).…”

Preparation and Characterization of Recombinant Dolphin Fish (Coryphaena hippurus) Growth Hormone

“…This latter method does not provide information on whether the purified hormone exists as a monomer or as a mixture of monomers, noncovalent dimers, or oligomers that cannot be identified by SDS-PAGE (11). In contrast, the reports from our laboratory regarding the preparation of three, to date prepared, fish GHs (12)(13)(14)(15) or other proteins (11,16 -22) are always accompanied by a determination of monomer content using a sensitive gel-filtration method. In many cases (12,13,16 -20), we have shown that only the monomers are biologically active despite the other forms' apparent purity on an SDS-polyacrylamide gel or their ability to interact with antibodies.…”

“…Preparation of the liver microsomal fraction from livers of dolphin fish and gilthead sea bream (Sparus aurata) and the binding experiments were carried out as described previously for common carp liver (12). To remove the endogenous GH, the microsomal fraction was desaturated with 4 M MgCl 2 as described previously (38).…”

Preparation and comparison of biological properties of recombinant carp (Cyprinus carpio) growth hormone and its Cys-123 to Ala mutant

“…Carp growth hormone (cGH) has also been cloned (Koren et al 1989;Chao et al 1987) and, more recently, expressed in E. coli as an insoluble aggregate (refractile bodies) (Fine et al 1993). Expression in E. coli as an insoluble aggregate is commercially preferable because of high yield, simpler and cheaper purification procedures and resistance to intracellular bacterial proteases (Sugimoto et al 1991b).…”