Zhang Yuebo, Liang Jing, Zhang Longchao, Wang Ligang, Liu Xin, Yan Hua, Zhao Kebin, Shi Huibi, Zhang Tian, Li Na, Pu Lei, Wang Lixian
Key Laboratory of Farm Animal Genetic Resources and Germplasm Innovation, Ministry of Agriculture, Institute of Animal Science, Chinese Academy of Agricultural Sciences, Beijing, 100193 China.
Jilin Academy of Agricultural Sciences, Changchun, 130033 China.
J Anim Sci Biotechnol. 2015 Dec 30;6:60. doi: 10.1186/s40104-015-0060-x. eCollection 2015.
In Sus scrofa, methionine sulfoxide reductase B3 (MSRB3) is a crucial candidate gene for ear size, and an important conformational trait of pig breeds. However, challenges in MSRB3 cDNA amplification have prevented further identification of MSRB3 allelic variants influencing pig ear size.
We cloned a full-length cDNA sequence of porcine MSRB3 by rapid-amplification of cDNA ends. The 3,765-bp gene contained a 5'-untranslated region (UTR) (190 bp), a coding region (552 bp), and a 3'-UTR (3,016 bp) and shared 84 %, 84 %, 87 %, 86 %, and 70 % sequence identities with human, orangutan, mouse, chicken, and zebrafish, respectively. The gene encoded a 183-amino acid protein, which shared 88 %, 91 %, 89 %, 86 %, and 67 % identities with human, orangutan, mouse, chicken, and zebrafish, respectively. Tissue expression analysis using qRT-PCR revealed that MSRB3 was expressed in the heart, liver, lung, kidney, spleen, ear, muscle, fat, lymph, skeletal, and hypothalamic tissues. Three single nucleotide polymorphisms (SNPs) were identified in MSRB3: c.-735C > T in the 5' flanking region, c.2571 T > C in the 3'-UTR, and a synonymous mutation of c.484 T > C in the coding region. The SNPs c.-735C > T and c.2571 T > C were significantly associated with ear size in a Large White × Minzhu F2 population other than in Beijing Black pigs. Subsequently, at SNP c.-735C > T, the mRNA of MSRB3 was significantly higher expressed in ears of individuals with the TT genotype (Minzhu) than those with CC (Large White).
The porcine MSRB3 owned a 3,765-bp full-length cDNA sequence and was detected to express in ear tissue. Two SNPs of this gene were shown to be significantly associated with ear size in a Large White × Minzhu intercross population instead of Beijing Black pig population. What's more, the individuals with higher mRNA expression of MSRB3 have larger ear sizes. These results provide useful information for further functional analyses of MSRB3 influencing ear size in pigs.
在野猪中，蛋氨酸亚砜还原酶B3（MSRB3）是耳部大小的关键候选基因，也是猪品种的一个重要形态特征。然而，MSRB3 cDNA扩增方面的挑战阻碍了对影响猪耳部大小的MSRB3等位基因变异的进一步鉴定。
我们通过cDNA末端快速扩增克隆了猪MSRB3的全长cDNA序列。这个3765 bp的基因包含一个5'非翻译区（UTR）（190 bp）、一个编码区（552 bp）和一个3'UTR（3016 bp），与人、猩猩、小鼠、鸡和斑马鱼的序列同一性分别为84%、84%、87%、86%和70%。该基因编码一个183个氨基酸的蛋白质，与人、猩猩、小鼠、鸡和斑马鱼的同一性分别为88%、91%、89%、86%和67%。使用qRT-PCR进行的组织表达分析显示，MSRB3在心、肝、肺、肾、脾、耳、肌肉、脂肪、淋巴、骨骼和下丘脑组织中表达。在MSRB3中鉴定出三个单核苷酸多态性（SNP）：5'侧翼区的c.-735C>T、3'UTR中的c.2571 T>C以及编码区的同义突变c.484 T>C。除北京黑猪外，SNP c.-735C>T和c.2571 T>C在大白猪×民猪F2群体中与耳部大小显著相关。随后，在SNP c.-735C>T处，MSRB3的mRNA在TT基因型（民猪）个体的耳中表达显著高于CC基因型（大白猪）个体。
猪MSRB3拥有一个3765 bp的全长cDNA序列，且在耳部组织中被检测到表达。该基因的两个SNP在大白猪×民猪杂交群体而非北京黑猪群体中与耳部大小显著相关。此外，MSRB3 mRNA表达较高的个体耳部较大。这些结果为进一步分析MSRB3对猪耳部大小的功能提供了有用信息。
