Reference : Production and purification of biologically active recombinant tilapia (Oreochromis nilo...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
http://hdl.handle.net/2268/20092
Production and purification of biologically active recombinant tilapia (Oreochromis niloticus) prolactins
English
Swennen, D. [> > > >]
Rentier-Delrue, Françoise mailto [Université de Liège - ULg > Département des sciences de la vie > Biologie et génétique moléculaire - GIGA-R : Coordination scientifique >]
Auperin, B. [> > > >]
Prunet, P. [> > > >]
Flik, G. [> > > >]
Wendelaar Bonga, S. E. [> > > >]
Lion, Michelle mailto [Université de Liège - ULg > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]
Martial, Joseph mailto [Université de Liège - ULg > Département des sciences de la vie > GIGA-R : Biologie et génétique moléculaire >]
1991
Journal of Endocrinology
Society for Endocrinology
131
2
219-27
Yes (verified by ORBi)
0022-0795
1479-6805
Bristol
United Kingdom
[en] Animals ; Blotting, Western ; Electrophoresis, Polyacrylamide Gel ; Fishes/*metabolism ; Kidney/metabolism ; Plasmids ; Prolactin/*biosynthesis/isolation & purification/metabolism ; Protein Binding ; Protein Engineering/methods ; Recombinant Proteins/*biosynthesis/isolation & purification/metabolism
[en] Recombinant expression vectors carrying tilapia prolactin-I or -II (tiPRL-I or tiPRL-II) cDNA were constructed and the tiPRL-I and II proteins were produced in E. coli as inclusion bodies. These inclusion bodies were dissolved in 6 mol urea/l. Refolding of the proteins was followed by SDS-PAGE under non-reducing conditions so as to visualize the oxidized state of the molecules. Proteins tiPRL-I and tiPRL-II were purified by gel filtration and ion-exchange chromatography. The N-terminal sequence and bioactivities of both purified proteins were then analysed. Recombinant tiPRL-I and tiPRL-II induced a significant rise in plasma calcium levels as well as in mucocyte density in the abdominal skin epithelium. When tested on kidney membrane, both proteins exhibited potency in competing with 125I-labelled tiPRL-I for binding sites, but tiPRL-I seemed to be more potent than tiPRL-II in competing for these sites. The results obtained for the biological activities tested suggest that both recombinant prolactins were correctly refolded and had retained the full biological activity previously observed with the natural hormone preparations extracted from the animals.
http://hdl.handle.net/2268/20092
http://joe.endocrinology-journals.org/cgi/content/abstract/131/2/219
1991/11/01

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