[Identification and analysis of a novel microsatellite marker flanking porcine myostatin gene (MSTN)].
In animal breeding, microsatellite marker plays an important role in constructing genetic maps, QTL mapping and function analysis of structural genes. Myostatin, also known as GDF8, is a negative regulator of skeletal muscle mass and, in swine, it is evidenced to be related to birth weight and average daily gain from 60 kg to 100 kg of body weight. In present study, by subcloning and sequencing,we identified a novel microsatellite marker which is useful for fine QTL mapping for meat traits. A BAC clone containing porcine MSTN was extracted and digested with EcoR I to recover the fragment of > 4 kb for subcloning in pGEM-3zf (+). Sequencing and alignment results showed that this subcloned fragment was not from porcine MSTN, but included a tandem repeat of (TG) 13, which is a novel microsatellite marker (GenBank accession number: AF454400) flanking MSTN. To exclude its vector origin we designed specific primers flanking this marker and successfully amplified this fragment from porcine genome. Through a pedigree analysis of a double-muscled Yorshire strain, we found that it is inherited in a co-dominant manner. We also checked the gene frequencies of this locus in 381 unrelated individuals of 7 pig breeds, namely Laiwu,Landrace, Yorkshire,Duroc, Peterian, Min and Erhualian. Only two alleles were detected, the repeating number of which are 13 (allele A) and 19 (allele B) respectively, which indicated that it is a low poly morphic microsatellite marker. In addition, the frequencies of the two alleles are different between the two types of pig breeds, while allele A is dominant in Chinese local breeds, allele B is dominant in imported breeds. Alignment with AY208121 indicate that this locus is located 42 kb downstream of porcine MSTN. We speculate that this microsatellite DNA is an important marker both in fine QTL mapping for meat traits and in the expression study of porcine MSTN.
National Lab for Agrobiotechnology, China Agricultural University, Beijing 100094, China.
Yun-Liang
YL
Jiang
Ning
N
Li
Xing-Bo
XB
Zhao
Xiao-Xiang
XX
Hu
Zhao-Liang
ZL
Liu
Xue-Mei
XM
Deng
Chang-Xin
CX
Wu
Li-Xin
LX
Du
Ji-Sheng
JS
Cao
D000818:Animals; D001483:Base Sequence; D022202:Chromosomes, Artificial, Bacterial; D005787:Gene Frequency; D018895:Microsatellite Repeats; D008969:Molecular Sequence Data; D055435:Myostatin; D013552:Swine / Q000235:genetics*; D016212:Transforming Growth Factor beta / Q000235:genetics*
D055435:Myostatin; D016212:Transforming Growth Factor beta