Differential expression profiling of ovarian genes in prelaying and laying geese
Differential expression profiling of ovarian genes in prelaying and laying geese
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DOI：
10.3382/ps.2008-00519
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发表时间：
2009-09-01
期刊：
影响因子：
4.4
通讯作者：
Dong, C. Y.
中科院分区：
文献类型：
--
作者：
Kang, B.;Guo, J. R.;Dong, C. Y.
鉴定产蛋鹅和预产鹅卵巢中基因的差异表达，是提高产蛋鹅产蛋性能的必要条件。本研究以卵巢为模型，采用抑制减法杂交和反向斑点杂交技术对这些基因进行鉴定。利用实时荧光定量PCR技术分析了鹅卵巢中雌激素受体1、雌激素受体2、促卵泡激素受体、催乳素受体、铁蛋白H链和卵巢差异表达未知基因08的表达谱。取产仔鹅和预产仔鹅子房总RNA并进行分离。从产蛋鹅分离的cDNA中减去产蛋鹅卵巢mRNA反转录的cDNA。通过反向斑点杂交进一步鉴定了含有推定差异表达基因片段的465个克隆。对97个克隆进行测序和进一步分析。序列分析表明，产蛋鹅卵巢中有18个已知基因片段（包括一个线粒体基因）和8个未知基因片段的表达量高于预产鹅。已知的17个基因编码的蛋白质属于与结合、催化活性、酶调节活性、信号换能器活性、结构分子和转运蛋白活性相关的组。实时荧光定量PCR结果显示，产蛋鹅卵巢中雌激素受体1、雌激素受体2、促卵泡激素受体、催乳素受体、铁蛋白H链和卵巢差异表达未知基因08的表达量均高于预产鹅（P < 0.05）。这些差异表达基因可能与产蛋鹅向产蛋阶段的发育有关。为进一步阐明鹅产蛋的分子机制，提高产蛋鹅的生产能力，还需要进一步的研究。
The identification of the differential expression of genes in the ovaries of egg-laying and prelaying Zi geese is required to improve the laying performance of the geese. In the present study, suppression subtractive hybridization and reverse dot-blot were employed to identify such genes, using the ovary as a model. Furthermore, expression profiling of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 in ovaries from geese was performed by quantitative real-time PCR. Total RNA from the ovaries of laying and prelaying Zi geese was pooled and the mRNA was isolated. The cDNA that was reverse-transcribed from the ovarian mRNA of the prelaying geese was subtracted from the cDNA isolated from the laying geese. Four hundred sixty-five clones containing putative differentially expressed gene fragments were further identified by reverse dot-blot. Ninety-seven clones were subjected to sequencing and further analysis. Sequence analysis showed that the expression of 18 known (including a mitochondrial gene) and 8 unknown gene fragments was higher in the ovaries of laying geese compared with prelaying geese. Seventeen of the known genes encode proteins that belong to groups involved with binding, catalytic activity, enzyme regulatory activity, signal transducer activity, structural molecule, and transporter activity. The results of the quantitative real-time PCR showed that the expression of estrogen receptor 1, estrogen receptor 2, follicle stimulating hormone receptor, prolactin receptor, ferritin H chain, and ovary differentially expressed unknown gene 08 was higher in the ovaries of the laying geese than in those of the prelaying geese (P < 0.05). These differentially expressed genes may be relevant to the progression of prelaying geese to the egg-laying stage. Further study is required to elucidate the molecular mechanism that controls egg-laying in geese, to improve the productivity of laying geese.