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Isoelectric focusing in immobilized pH gradients with carrier ampholytes added for high‐resolution phenotyping of bovine β‐lactoglobulins: Characterization of a new genetic variant
Author(s) -
Krause Ingolf,
Buchberger Johann,
Weiß Günther,
Pflügler Monika,
Klostermeyer Henning
Publication year - 1988
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150090925
Subject(s) - isoelectric focusing , chemistry , isoelectric point , immobilized ph gradient , chromatography , resolution (logic) , genetic variants , amino acid , analytical chemistry (journal) , biochemistry , enzyme , artificial intelligence , computer science , genotype , gene
The genetic variants of bovine β‐lactoglobulin (β‐lg) from the “Murnau‐Werdenfelser” breed were analyzed in three different isoelectric focusing (IEF) systems. While carrier ampholyte IEF with a pH gradient of 0.2 pH/cm did not resolve the new variant W from the B variant and IEF in immobilized pH gradients (IPG) with 0.1 pH/cm only partially resolved it, adequate separation was achieved with IPG‐IEF in a pH 5.25‐pH 5.7 gradient, in presence of 0.8 % w/v carrier ampholytes, both over a 10 and 17 cm separation distance. Apparent isoelectric points (p I 's) and genetic frequencies (f) were as follows: β‐lg A, p I = 5.370, f = 0.364; β‐lg B, p I = 5.485, f = 0.480; β‐lg W, p I = 5.492, f = 0.076; and β‐lg D, p I = 5.610, f = 0.080. The small difference of Δ p I = 0.007 between β‐lg B and β‐lg W respectively, seems to originate from a “silent” substitution of neutral amino acid residues as compared to the larger Δ p I 's of the other genetic variants of β‐lg, which result from substitution of charged amino acids.