One-step, non-denaturing purification method of carp (Cyprinus carpio) vitellogenin.
A new single-step purification procedure was developed to purify carp (Cyprinus carpio) vitellogenin (VTG), from estradiol-treated carp plasma. This method was performed by high performance liquid weak anion-exchange chromatography, using a discontinuous elution gradient of NaCl (0-0.5 M, steps of 12.5 mM/4 min). SDS and native-PAGE analysis, of treated-fish plasma and purified solution, showed the appearance of a 370 kDa phospholipoprotein, composed of two 130 kDa monomers, with all VTG characteristics. The sequencing of a 130 kDa monomer confirmed that it was carp VTG. Consequently, this procedure is a rapid method, permitting high quantities of non-denatured carp VTG to be obtained.
Magalhães
Ilizabete
I
Laboratoire d'Immunologie et de Microbiologie (LIM/ESE-CNRS, unité FRE2635), Dept. Genie Biologique, IUT de Thionville-Yutz, 1 Impasse A. Kastler, F-57970 Yutz, France.
Ledrich
Marie-Laure
ML
Pihan
Jean-Claude
JC
Falla
Jaïro
J
D000595:Amino Acid Sequence; D000818:Animals; D002347:Carps; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D002852:Chromatography, Ion Exchange / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D014819:Vitellogenins / Q000737:chemistry / Q000302:isolation & purification*
D014819:Vitellogenins