TY - JOUR
T1 - Identification of Sheep Ovary Genes Potentially Associated with Off-season Reproduction
AU - Chen, Lei
AU - Liu, Ka
AU - Zhao, Zongsheng
AU - Blair, Hugh T.
AU - Zhang, Peng
AU - Li, Daquan
AU - Ma, Runlin Z.
N1 - Funding Information:
This work was partially supported by the grants from the Ministry of Science and Technology of China (No. 2007DFB30420 ), and the China Ministry of Agriculture (No. 2009ZX08008-005B ). We thank Wengxiang Zhang and Songhua Zhao for assistance in observation and time-recording of the sheep oestrus in experimental herds.
PY - 2012/4/20
Y1 - 2012/4/20
N2 - Off-season reproduction is a favorable economic trait for sheep industry. Hu sheep, an indigenous Chinese sheep breed, demonstrates a higher productivity of lambs and displays year-around oestrous behavior under proper nutrition and environment. The genetic basis behind these traits, however, is not well understood. In order to identify genes associated with the off-season reproduction, we constructed a suppression subtractive hybridization (SSH) cDNA library using pooled ovary mRNAs of 6 oestrous Hu females as a tester and the pooled ovary mRNAs of 6 non-oestrous Chinese Merino females as a driver. A total of 382 resulting positive clones were obtained after the SSH. We identified 114 differentially up-regulated genes in oestrous Hu sheep by using subsequent screening and DNA sequencing, of which 8 were previously known, 93 were reported for the first time in sheep, and 13 were novel with no significant homology to any sequence in the DNA databases. Functions of the genes identified are related to cell division, signal transduction, structure, metabolism, or cell defense. To validate the results of SSH, 6 genes (Ntrk2, Ppap2b, Htra1, Nid1, Serpine2 and Foxola) were selected for conformational analysis using quantitative real-time PCR (qRT-PCR), and two of them (Htral and Foxo1a) were verified by Northern blot. All of the 6 genes were differentially up-regulated in the ovary of oestrous Hu. It is obvious that off-season reproduction is a complex trait involving multiple genes in multiple organs. This study helps to provide a foundation for the final identification of functional genes involved in the sheep ovary.
AB - Off-season reproduction is a favorable economic trait for sheep industry. Hu sheep, an indigenous Chinese sheep breed, demonstrates a higher productivity of lambs and displays year-around oestrous behavior under proper nutrition and environment. The genetic basis behind these traits, however, is not well understood. In order to identify genes associated with the off-season reproduction, we constructed a suppression subtractive hybridization (SSH) cDNA library using pooled ovary mRNAs of 6 oestrous Hu females as a tester and the pooled ovary mRNAs of 6 non-oestrous Chinese Merino females as a driver. A total of 382 resulting positive clones were obtained after the SSH. We identified 114 differentially up-regulated genes in oestrous Hu sheep by using subsequent screening and DNA sequencing, of which 8 were previously known, 93 were reported for the first time in sheep, and 13 were novel with no significant homology to any sequence in the DNA databases. Functions of the genes identified are related to cell division, signal transduction, structure, metabolism, or cell defense. To validate the results of SSH, 6 genes (Ntrk2, Ppap2b, Htra1, Nid1, Serpine2 and Foxola) were selected for conformational analysis using quantitative real-time PCR (qRT-PCR), and two of them (Htral and Foxo1a) were verified by Northern blot. All of the 6 genes were differentially up-regulated in the ovary of oestrous Hu. It is obvious that off-season reproduction is a complex trait involving multiple genes in multiple organs. This study helps to provide a foundation for the final identification of functional genes involved in the sheep ovary.
KW - CDNA library
KW - Hu sheep
KW - Oestrous
KW - Ovary
KW - Suppression subtractive hybridization
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U2 - 10.1016/j.jgg.2012.03.002
DO - 10.1016/j.jgg.2012.03.002
M3 - Article
C2 - 22546540
AN - SCOPUS:84860380757
SN - 1673-8527
VL - 39
SP - 181
EP - 190
JO - Journal of Genetics and Genomics
JF - Journal of Genetics and Genomics
IS - 4
ER -