1022-7954/04/4009- © 2004
Russian Journal of Genetics, Vol. 40, No. 9, 2004, pp. 1027–1030. Translated from Genetika, Vol. 40, No. 9, 2004, pp. 1251–1255.
Original Russian Text Copyright © 2004 by Ludvikova, Dzisuk, Aytkhozhina.
Rapid accumulation of the data on mitochondrial
genome polymorphism in human populations along
with speciﬁc features of mtDNA, including high muta-
tion rate, lack of recombinations, and maternal trans-
mission, provided identiﬁcation of ethnic, racial and
population-speciﬁc polymorphic sites .
In population respect, restriction polymorphism
within the main noncoding region of mtDNA and dele-
tion/insertion polymorphism in the noncoding inter-
genic region, located between the cytochrome oxidase
II and lysine tRNA genes, the so-called V region, are
most thoroughly studied [2–4]. The latter segment
includes two tandem copies of the nine-nucleotide
sequence, CCCCCTCTA . The deletion of one of
these copies is considered typical of the populations of
Asia and Oceania [6–8]. Much rarer mutation in this
region is the insertion of four cytosines, CCCC [9–11].
Correlation analysis carried out using the combined
data for populations of Siberia, Asia, and Oceania has
revealed a geographic gradient of the deletion fre-
quency, which increased southward, and to a lower
extend, eastward [5, 8–10]. The trend azimuth of the
deletion frequency (
%) within the range from subarc-
tic regions of Siberia to western regions of Oceania is
almost completely described by a multiple regression
= 30.516 – 0.840
, geographic longitude, and
expected deletion value, calculated for particular terri-
tory). The proportion of geographic component of gen-
eral variability of the deletion frequency constitutes
64% . The pattern observed does not extend to
American Indians [11, 12]. Similar polymorphism was
described among the indigenous populations of Africa;
however, in this case the deletion had an independent
The distribution pattern of the region V deletion
among the Mongoloids with the maximum in Southeast
Asia and Oceania and with the minimum, up to the
complete absence, in Northeast Asia, is thought to be
associated with the settling patterns of ancient popula-
tions in Asia [9, 10]. Despite ample data on the deletion
frequency in the populations of Asia, precise determi-
nation of its distribution pattern in Asia requires addi-
tional investigation of this polymorphism in the previ-
ously not examined populations. There is also marked
uncertainty about the origin and the distribution pat-
terns of the V-region insertion in human populations.
For these reasons, the present study focused on the
analysis of insertion–deletion polymorphism of the
mtDNA V region in three populations of Kazakhs from
different regions of Kazakhstan.
MATERIALS AND METHODS
Total DNA samples were obtained from blood leu-
kocytes of Kazakh individuals from different regions of
Kazakhstan and belonging to three different contempo-
rary populations (Almaty, Semipalatinsk, and Altai).
DNA was isolated from the whole blood according to a
standard method [14, 15]. In accordance with the Cam-
bridge reference sequence, the primers used for ampli-
ﬁcation of V region were 8196–8220, forward, and
8296–8316, reverse . The reaction mixture con-
tained 67 mM Tris–HCl (pH 8.8); 16.6 mM
5 mM MgCl
; 0.01% Tween 20; 0.15 mg/ml albumin;
g genomic DNA; 2 pmol of each primer; and
0.2 mM of each dNTP (dATP, dGTP, dCTP, dTTP).
After initial denaturation for 5 min at 95
C 1 unit od
Biotag DNA polymerase (Dialat, Russia) was added.
Thirty-three cycles of ampliﬁcation were performed.
Restriction was performed for 2 h at 37
C in a 50-
ﬁnal volume, containing 2 units of the enzyme; 10 mM
Tris–HCl (pH 7.4); 10 mM MgCl
; and 1 mM DTT.
of Mitochondrial DNA Region V in Kazakh Populations
from Different Regions of Kazakhstan
E. K. Ludvikova, N. V. Dzisuk, and N. A. Aytkhozhina
Aytkhozhin Institute of Molecular Biology and Biochemistry, Almaty, 480012 Kazakhstan;
fax: (7-3272) 92-19-47; e-mail: firstname.lastname@example.org
Received August 20, 2003
—Insertion–deletion polymorphism of the mitochondrial DNA (mtDNA) region V was examined in
three Kazakh populations inhabiting different regions of Kazakhstan. The 9-bp deletion was revealed in all three
populations examined. In Altai population the 4-bp insertion was also found. The presence of these polymor-
phic variants was conﬁrmed by DNA sequencing.