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Identification of differentially expressed genes in Mongolian sheep ovaries by suppression subtractive hybridization

文献类型： 外文期刊

作者： He, Xiaolong 1 ; Li, Bei 3 ; Wang, Feng 1 ; Tian, Chunying 1 ; Rong, Weiheng 1 ; Liu, Yongbin 1 ;

作者机构： 1.Inner Mongolia Acad Agr & Anim Husb Sci, Hohhot 010031, Peoples R China

2.Chinese Acad Sci, Inner Mongolia Prataculture Res Ctr, Hohhot 010031, Peoples R China

3.Inner Mongolia Agr Univ, Coll Anim Sci, Hohhot 010018, Peoples R China

关键词： Mongolian sheep;Suppression subtractive hybridization;Expressed sequence tags

期刊名称：ANIMAL REPRODUCTION SCIENCE （ 影响因子：2.145； 五年影响因子：2.281 ）

ISSN： 0378-4320

年卷期： 2012 年 133 卷 1-2 期

页码：

收录情况： SCI

摘要： Fecundity is an important trait in sheep. Because it is directly related to production costs and efficiency, it has great economic impact in sheep husbandry. Because Mongolian sheep are a longstanding, indigenous breed, they are genetically related to most other breeds of sheep in China. The study of genes related to reproductive traits is essential to improving the fecundity of Mongolian sheep. In the present study, suppression subtractive hybridization (SSH) was performed using forward and reverse nested primers on cDNA libraries from ovarian tissue of single-bearing (S) and biparous (B) Mongolian sheep (MS). This yielded 768 clones. The length of the inserted fragments ranged from 150 to 1000 bp. From these, dot blot hybridization followed by sequencing and homology blast search in GenBank resolved 373 differentially expressed clones, representing 185 gene sequences (homology >85% and length >200 bp), 10 expressed sequence tags (ESTs; homology >95% and length >100 bp), and 4 unknown ESTs. The analysis of the differentially expressed gene functions allowed these genes to be categorized into seven groups: cell/body or immune defense, metabolism, transportation, nucleic acid modification, cell development, signal transduction, and cell structure. Four differentially expressed genes, a disintegrin and metalloproteinase with thrombospondin motifs 1 (ADAMTS1), inhibitor of DNA binding 3 (ID3), bone morphogenetic protein 6 (BMP6), and integrin beta 1 (ITGB1), were randomly selected and verified using relative quantitative real-time polymerase chain reaction (RQ-PCR). The expression of these genes in BMS ovaries was 30.06, 11.55, 0.82, and 1.12-fold that of SMS ovaries, respectively. (C) 2012 Elsevier B.V. All rights reserved.

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