Production and purification of biologically active recombinant tilapia (Oreochromis niloticus) prolactins. 1991

D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
Laboratoire de Biologie Moleculaire et de Génie Génétique, Université de Liège, Sart Tilman, Belgium.

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.

UI MeSH Term Description Entries
D007668 Kidney Body organ that filters blood for the secretion of URINE and that regulates ion concentrations. Kidneys
D010957 Plasmids Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS. Episomes,Episome,Plasmid
D011388 Prolactin A lactogenic hormone secreted by the adenohypophysis (PITUITARY GLAND, ANTERIOR). It is a polypeptide of approximately 23 kD. Besides its major action on lactation, in some species prolactin exerts effects on reproduction, maternal behavior, fat metabolism, immunomodulation and osmoregulation. Prolactin receptors are present in the mammary gland, hypothalamus, liver, ovary, testis, and prostate. Lactogenic Hormone, Pituitary,Mammotropic Hormone, Pituitary,Mammotropin,PRL (Prolactin),Hormone, Pituitary Lactogenic,Hormone, Pituitary Mammotropic,Pituitary Lactogenic Hormone,Pituitary Mammotropic Hormone
D011485 Protein Binding The process in which substances, either endogenous or exogenous, bind to proteins, peptides, enzymes, protein precursors, or allied compounds. Specific protein-binding measures are often used as assays in diagnostic assessments. Plasma Protein Binding Capacity,Binding, Protein
D011994 Recombinant Proteins Proteins prepared by recombinant DNA technology. Biosynthetic Protein,Biosynthetic Proteins,DNA Recombinant Proteins,Recombinant Protein,Proteins, Biosynthetic,Proteins, Recombinant DNA,DNA Proteins, Recombinant,Protein, Biosynthetic,Protein, Recombinant,Proteins, DNA Recombinant,Proteins, Recombinant,Recombinant DNA Proteins,Recombinant Proteins, DNA
D004591 Electrophoresis, Polyacrylamide Gel Electrophoresis in which a polyacrylamide gel is used as the diffusion medium. Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D005399 Fishes A group of cold-blooded, aquatic vertebrates having gills, fins, a cartilaginous or bony endoskeleton, and elongated bodies covered with scales.
D000818 Animals Unicellular or multicellular, heterotrophic organisms, that have sensation and the power of voluntary movement. Under the older five kingdom paradigm, Animalia was one of the kingdoms. Under the modern three domain model, Animalia represents one of the many groups in the domain EUKARYOTA. Animal,Metazoa,Animalia
D015153 Blotting, Western Identification of proteins or peptides that have been electrophoretically separated by blot transferring from the electrophoresis gel to strips of nitrocellulose paper, followed by labeling with antibody probes. Immunoblotting, Western,Western Blotting,Western Immunoblotting,Blot, Western,Immunoblot, Western,Western Blot,Western Immunoblot,Blots, Western,Blottings, Western,Immunoblots, Western,Immunoblottings, Western,Western Blots,Western Blottings,Western Immunoblots,Western Immunoblottings
D015202 Protein Engineering Procedures by which protein structure and function are changed or created in vitro by altering existing or synthesizing new structural genes that direct the synthesis of proteins with sought-after properties. Such procedures may include the design of MOLECULAR MODELS of proteins using COMPUTER GRAPHICS or other molecular modeling techniques; site-specific mutagenesis (MUTAGENESIS, SITE-SPECIFIC) of existing genes; and DIRECTED MOLECULAR EVOLUTION techniques to create new genes. Genetic Engineering of Proteins,Genetic Engineering, Protein,Proteins, Genetic Engineering,Engineering, Protein,Engineering, Protein Genetic,Protein Genetic Engineering

Related Publications

D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
December 1993, General and comparative endocrinology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
May 2019, Fish & shellfish immunology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
February 1994, Journal of molecular endocrinology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
October 1997, General and comparative endocrinology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
October 1995, Archives of biochemistry and biophysics,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
January 2012, Applied biochemistry and biotechnology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
August 1992, General and comparative endocrinology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
August 2017, Veterinary microbiology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
April 1994, The American journal of physiology,
D Swennen, and F Rentier-Delrue, and B Auperin, and P Prunet, and G Flik, and S E Wendelaar Bonga, and M Lion, and J A Martial
October 1995, Biochimica et biophysica acta,
Copied contents to your clipboard!