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cattle br breeds. were cle subjecte showed Bali catt haplotyp indicated the cattl polymorKey wo
INTROD
Huitema
tropical
have fo
(native
11
GENET
Muh
esearch Cen
boratory of B
gyakarta, Ind
adjah Mada
RACT
The aim of t ome b (cyt us). This reg
on fragment reed as sam
The L14735 eaved by Taq ed to diterm
that Indone tle, Madura c pe A and B
d that the ca e related to rphism of
mt-ords: PCR-R
DUCTION
The develo
a (1982) re
environme
our domest
Pacitan cat
TIC POLYM
IN
ammadCa
nter for Biot
Biochemistr
donesia; La
a University
this research b) mitochon
gion is wide length polym mples, there 5 and H1514 qI restriction
ine the dive esian domes
cattle and Lo , and one o attle related Bos indicus
-DNA cytoch
RFLP, mtDNA
oping of dom
eported ther
ent and man
tic cattle br
ttle).
MORPHISM
INDONESI
ahyadi1,3, W
technology,
ry, Faculty o
aboratory o
y, Yogyakart
h was to iden ndrial gene ely used as morphism (PC e were Bali, 49 primer pai
enzyme and ersity of dom
tic cattle ha ocal cattle of of Local catt to Bos javan s. Indonesian
rome b.
A cyt b, Indo
mestic cattle
re domestic
nagement, a
reed, such
OF mt-DN
IAN DOMES
WayanT. Ar
, Gadjah Ma
of Veterinar
f Animal Br
ta, Indones
ntify nucleotid of Indonesi a target po CR-RFLP) fo Madura, O ir was used d then electro mestic cattle
d three hap Pacitan inclu tle of Pacita nicus (Bante
n domestic c
onesian Dom
e is very im
c cattle has
and more re
as Bali, M
NA CYTOCH
STIC CATT
rtama1,2 an
ada Univers
ry Medicine
reeding, Fac
ia (contact
de polymorph ian Domesti olymorphism or species ide
ngole-Grade to amplify th ophoreted on
base on m lotype. The uded in haplo an belong to eng), while H
cattle had ge
mestic Cattle,
mportant for
s better rep
esistant from
Madura, PO
HROME B (
TLE
d Tety Hart
sity, Yogyak
e, Gadjah M
culty of Anim
author: tety
hic sites in a ic Cattle (B by polyme entification s e (PO) and he cyt b gene n 2% agaros maternal inhe
domestic ca otype A, the o haplotype Haplotype B enetic divers
Haplotype
Indonesian
roductivity,
m tropical d
O and Loca
(CYT B)
tatik3
karta, Indon
ada Univer
mal Husban
a 464 bp regi os indicus
rase chain studies. We u
Local Pacita e. The PCR
e gels. This eritance. The attle which c
PO cattle inc C. The hap and C indic sity based on
n livestock i
more adap
disease. Ind
al cattle of products research e results consist of cluded in plotype A ated that n genetic
ndustry.
ptable in
donesian
the seq
from Ma
NTB; B
were pr
appropr
K extrac
was car
PCR bu
TGT TA
univers
polyme
pre-den
final ex
examin
ethidium
by restr
a 2% ag
RESUL
Polymo
was res
To study th
quence of m
n. Mitochon
s much fas
red to nucle
96). In add
cygosity. Th
k and Grzy
als, were
enetic studie
RIALS AND
Samples c
adura Islan
ali cattle). T
repared by
riate with it’
ction (Samb
PCR and R
naturation, 3
xtension ste
ed by elect
m bromide.
riction enzy
garose gel.
LTS AND D
orphism of
Polymerase
sulted 464 b
he genetic
mt-DNA cyto
ndrial DNA
ster than nu
ear DNA, fa
dition, mate
he one of ge
ybowski, 2
mainly inv
es (Wolf et
D METHODS
collection a
d, Pacitan R
The DNA w
using DNA
’s protocol.
brook et al.,
RFLP anal
a final volu
MgCl2, 0.1
CT A and
ernal prime
µl DNA gen
35 cycles o
ep of 3 mi
trophoresis
As size ref
mes as des
In this work
ISCUSSION
f mt-DNA C
e chain rea
bp. The PCR
polymorphi
ochrome b i
had been
uclear (nc)
acilitating th
ernal inheri
ene that en
004). Cyto
vestigated f
al., 1999).
S
and DNA ex
Regency, Y
was isolated
A isolation
Ear tissue
, 1989).
ysis. Ampl
ume of 20 µ
1 µl dNTP m
H15149: G
r pair, desig
nom. The c
of 36 s at 9
n at 72oC
through a
ference, No
scribed prev
k we used T
NS
Cytochrome
action was c
R product w
ism of dom
n several b
used as a
DNA and
he identifica
tance of th
ncoded by m
chrome b
for evolutio
xtraction.
Yogyakarta
d from blood
KIT high pu
s were prep
lification of
l in 0.5 ml t
mix, 1 µl ea
GCC CCT C
gned by Ko
cycling cond
95oC, 73 s a
and 4oC u
1% agaros
vagen mark
viously (Ver
TaqI restrict
e b based o
carried out i
was digeste
mestic cattle
reeds which
a molecula
thus conta
ation of clos
he mt-DNA
mitochondri
gene of s
onary, gen
Both blood
and anothe
d samples a
ure PCR te
pared by us
the mt-DN
tubes conta
ach primer (
CAG AAT
ocher et al.
ditions were
at 51oC, 84
until the ne
se gel in 10
ker was use
rkaar et al.,
tion enzyme
on TaqI-RF
in a final vo
d by TaqI re
es in Indone
h collected
r marker. M
ains more s
sely related
A generally
al DNA is c
several ver
etic divers
and ear tis
er region (Ba
and ear tiss
emplate pre
sing standa
NA cytochro
aining 13.3
(L14735: AA
GAT ATT
(1989), 0,5
e as follows
4 s at 72oC
FLP Analys
olume 20 µl
estriction en
esia, we ex
from the dif
Mitochondr
sequence d
species (B
results in
cytochrome
rtebrates, in
sity and m
ssue were c
ali, Kaliman
sue. Blood s
eparation (R
ard SDS/pro
ome b (cyt
and follow
CR produc
ffer and sta
oduct was d
electrophor
sis
. The PCR
nzyme (Fig
xamined
fference
ial DNA
diversity
Brown et
lack of
e b gene
ncluding
molecular
collected
ntan and
samples
ROCHE)
oteinase
b) gene
digested
reted on
product
Figure Bali cat cattl
Native
cattle f
especia
Sequen
from NC
that PO
NC0059
taurus c
RFLP p
(haploty
416 and
e 1. The TaqI ttle (similar p e (Pc11), lan
Figure 1 s
s were grou
48 bp (lane
haplotype
pe B. The
ces analys
Pacitan ca
from DIY (
ally Pc11 ca
nces Analy
Three diffe
CBI. Seque
O cattle (PO
971. This r
cattle. Prad
pattern and
ype B).
One of nati
d 48 bp. it’s
-RFLP analy pattern with M ne 3 is PO ca
show Taq
I-uped at thre
e 1), was h
C; and the
e cleaved
sis with DN
ttle include
Bos indicu
attle include
ysis of Thre
erent individ
ences analy
O22) have s
results indi
o et al (200
d produced
ive Pacitan
s sequence
ysis of indone Madura and P attle from DIY ne
RFLP patte
ee different
haplotype A
third patter
fragment
NAMAN sof
d to the ha
us) included
ed to the hap
ee Differen
dual haploty
ysis was pe
similar Taq
cated that
05) reported
d three DN
cattle (Pc1
and TaqI-R
esian domes PO cattle fro Y (PO22) an egative contr
ern of Indo
t haplotype.
A; second pa
rn generate
sizes wer
ftware. The
aplotype A
d to the ha
plotype B.
t Haplotyp
ype sequen
erformed by
qI-RFLP pat
PO cattle
d that Bos in
NA band w
1) was hap
RFLP patter
stic cattle’s m om Pacitan R d lane 4 is th rol.
onesian do
. The first p
attern gene
d 372 bp, 4
re determin
e results sh
(the compl
aplotype C
pe in Indone
nces comp
DNAMAN
ttern with th
related to
ndicus and
ith 372 bp
plotype C. It
rn almost si
mt-DNA cytoc Regency), lan he PCR prod
omestic catt
pattern gene
erated 416 b
48 bp, and 4
ned by in
howed that
lete data n
C, while Lo
esian Dom
ared to the
software. T
he AF4923
Zebu (Bos
Bos taurus
, 48 bp an
t produced t
milar with t
chrome b. La ne 2 is Local duct (464 bp)
tle. The re
erated 225
bp and 48 b
44 bp (lane
dividual ha
Bali, Mad
ot shown),
ocal Pacitan
mestic Cattle
e some seq
The results
51, AF4923
s indicus) a Pacitan ) as a
estriction
bp, 191
bp (lane
e 3), was
aplotype
ura and
the PO
n cattle,
e
quences
showed
Bos Ta
number
NC0059
Figure box
included
lineage
betwee
cattle w
due to B
aurus sequ
r 93, CÆA
971).
e 2. Polymorp x indicated Ta
The Bali, M
d to the ha
(maternal
n Zebu and
was domest
Bali, Madur
uences. Th
A (Fig. 2,
phism of Indo aqI restrictio
Madura and
aplotype A.
inheritance
d Banteng c
ticated ban
ra and nativ
e differenti
compared
onesian Dom n site. The s
d some nat
. This resu
e). The Mad
cattle (Nijm
teng (Anon
ve Pacitan c
iation betw
lane Pc11
mestic cattle equences an
software.
tive Pacitan
lt indicated
dura cattle
an et al., 2
nimous, 200
cattle conta
ween them
1 with lane
sequences i nalysis was p
n (complete
d that the c
suspected
003; Wijono
05). The ge
in Banteng
due to alt
e AF49235
n three differ performed w
e data not s
cattle had t
as a result
o and Setia
enetic chara
nucleotides
teration nu
51, AF4923
rent haplotyp ith the DNAM
shown) catt
the same m
t of cross b
adi, 2004). T
acteristics s
s in it’s mt-D
ucleotide
350 and
pe. The MAN
tle were
maternal
breeding
The Bali
similarity
b gene.
with the
CONCL
based o
A, B an
Banteng
ACKNO
Mada U
the staf
REFER
. It supporte
e AY689188
LUSIONS
There wer
on TaqI-RF
nd C. Sequ
g nucleotide
OWLEDGE
We are tha
University fo
ff Veterinary
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