ELSEVIER
Journal of Chromatography A, 768 (1997) 47-56
JOURNALOF
CHROMATOGRAPHY A
Capillary electrophoretic analysis of genetic variants of milk
proteins from different species
Isidra Recio, Maria-Luisa Perez-Rodrlguez , Mercedes Ramos, Lourdes Amigo*
lnstituto de Fermentaciones lndustriales (CS1C), Juan de la Cierva 3, 28006 Madrid, Spain
Abstract
Polymorphism of bovine, ovine and caprine milk proteins was studied by CE. Identification of some rare bovine variants
was carried out by isoelectric focussing (IEF) using PhastSystem. Genetic variants A and D of bovine a 2-casein, [3-casein
variants A', A2, A 3, B and C and %~-casein variants B and C were determined by CE. In addition, the different casein
fractions including some genetic variants of ovine and caprine milk were identified by CE. In order to carry out this
identification, collected fractions from a cation-exchange FPLC separation were injected by CE.
Keywords: Milk; Proteins; Caseins
1. Introduction
Milk composition and the main characteristics of
its various constituents are now well known. In
particular the amino acid sequences of its seven main
protein components have been elucidated. At the
moment, there is no other food product whose
proteins are so well characterized. Moreover, for six
of these seven protein types a qualitative polymor-
phism has been described [1].
Genetic polymorphism in the milk proteins is due
to either substitutions of amino acids, or deletion of a
certain amino acid sequence along the peptide chain,
consequently these mutations are caused by changes
in the sequence of base pairs of the DNA molecule
which constitute the protein gene. It has been
described that polymorphism is also due to post-
transcriptional modifications, such as different de-
*Corresponding author.
'Present address: Facultad de Farmacia, Universidad Com-
plutense, 28040 Madrid, Spain.
grees of phosphorylation and glycosylation of the
protein. Besides the structural polymorphism, a
quantitative allelic variability has been found which
renders differences in protein expression. This phe-
nomenon has been described in cow [2] and goat
milk [3].
Polymorphism of milk proteins is related to pro-
duction traits, milk composition and milk quality.
Rapid screening of milk protein variants is important
for the genetic improvement of milk with regard to
the potential technological application and breeding
selection programs [4].
Until now, the detection of genetic polymorphism
of milk proteins has been achieved through various
electrophoretic and chromatographic techniques [1].
However, the conventional electrophoretic methods
are restricted to the resolution of proteins with
differing net charges caused by the substitution of
one amino acid for another and it is only possible to
achieve semiquantitative results. Peptide mapping by
HPLC and DNA mapping have also been used for
the detection of silent variants (mutations in proteins
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