ISSN 1022-7954, Russian Journal of Genetics, 2009, Vol. 45, No. 12, pp. 1454–1457. © Pleiades Publishing, Inc., 2009.
The myostatin (
) gene has been considered as
an important candidate gene for growth and develop-
ment of domestic animals because of its key role in
muscle growth and its potential applications in animal
husbandry. Many studies have focused on the correla-
tion between the polymorphism of
gene and the
growth traits in pig [1–3], cattle [4, 5] and chicken
[6, 7]. In goat, Li et al.  identiﬁed eight single-nucle-
otide polymorphisms of the
gene by sequencing
and found four haplotypes. The sheep
been sequenced for promoter region (AY918121),
intron 1 (AF393618), exon 2 and exon 3 (AF266758).
However, the polymorphism of sheep
not been reported so far. In this study, part of
gene from sixty sheep belonging to nine Chinese indig-
enous sheep breeds and one foreign breed were
sequenced in order to provide bioinformatic informa-
gene in the Ovis aries species which will
help in studying of
gene in domestic animal.
MATERIALS AND METHODS
. A total of sixty ani-
mals of ten sheep breeds were sampled from seven
The article is published in the original.
provinces in China. Of the ten breeds, nine were Chi-
nese indigenous breeds (Tan sheep, Hu sheep, Small
Tailed Han sheep, Ujumqin sheep, Hazake sheep, Altay
Fat-rumped sheep, Tong sheep, Tibetan sheep and
Monggol sheep) and one imported breed (Poll Dorset).
The sampling places, sample sizes, sample type, and
breed type of each sheep breed were given in Table 1.
The DNA was extracted from blood or kidney by the
phenol extraction method. The quality of DNA was
checked on 0.8% agarose and the quantity was checked
by UV spectrophotometer at A260/A280 nm. The sam-
ples having OD ratio between 1.7–1.9 were considered
good, and used for polymerase chain reaction.
Primer design, PCR ampliﬁcation and sequencing.
The 378-bp fragment of
gene was ampliﬁed with
a pair of primers designed using Primer3
based on sheep
sequence (AF266758). The for-
ward and reverse primers were 5'-ccctccctttactgtcatcc-3'
and 5'-atcaagcccaaaatctctcc-3', respectively, and they
were synthesized by Shanghai Sangon Biological Engi-
neering & Technology services Co., Ltd. (Shanghai,
China). The fragment ampliﬁed spanned bases from
1917 to 2294 including intron2 and exon3.
The polymerase chain reaction was carried out in
PTC-100TM Programmable Thermal Controller
SNP Detection and Haplotype Analysis in Partial
Sequence of MSTN Gene in Sheep
Y. F. Gong
, X. L. Li
, Z. Z. Liu
, X. M. Jin
, R. Y. Zhou
, L. H. Li
, and Q. Zhang
Hebei Key Laboratory of Veterinary Preventive Medicine, Department of Animal Science,
Hebei Normal University of Science & Technology, Changli, 066600, China
Department of Animal Science, College of Animal Science and Technology,
Hebei Agricultural University, Baoding, 071001, China;
Key Laboratory Animal Genetics and Breeding of the Ministry of Agricultural, State Key Laboratory of AgroBiotechnology,
College of Animal Science and Technology, China Agricultural University, Beijing 100094, China
Received June 18, 2008; in ﬁnal form, April 01, 2009
—Polymerase chain reaction (PCR) products of the
gene ampliﬁed from sixty sheep of nine
Chinese indigenous sheep breeds and one imported sheep breed were sequenced to identify the single-nucle-
otide polymorphisms (SNPs) in a 378-bp fragment including intron 2 and exon 3 of the
gene. A total of
ﬁfteen SNPs (A1937C, T1942G, C1956T, A1972C, A1990G, A2008C, A2011G, C2019T, A2025C, A2027C,
T2085G, T2173C, C2198T, C2210T and C2213T) were detected among the sixty sequenced individuals and
they were all located in intron 2. Twelve haplotypes were identiﬁed from these ﬁfteen SNPs, of which haplotype
I (CGTCGCGTCCGCTTT) and VIII (ATCAAAACAATTCCC) were the two major and basic ones with fre-
quencies of 12.25% and 77.80%, respectively. Haplotype VIII was distributed in all sheep breeds and all indi-
viduals of the meat or meat-wool type sheep breeds were homozygous with respect to this haplotype. This sug-
gests that haplotype VIII might be related to meat production traits in sheep. Haplotype I was only distributed
in the fur, lambskin type and fur-meat type sheep breeds. This suggests that haplotype I may have some rela-
tionship with fur traits in sheep.