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Molecular characterization, expression profile and transcriptional regulation of the CYP19 gene in goose ovarian follicles
Gene ( IF 3.5 ) Pub Date : 2021-08-26 , DOI: 10.1016/j.gene.2021.145928
Yan Deng 1 , Huilan Huang 1 , Yujing Rong 1 , Shenqiang Hu 1 , Jiwei Hu 1 , Bo Hu 1 , Hua He 1 , Liang Li 1 , Jiwen Wang 1
Affiliation  

Cytochrome P450 Family 19 (CYP19) is a crucial enzyme to catalyze the conversion of androgens to estrogens. However, the regulatory mechanism of goose CYP19 gene remains poorly understood. The present study attempted to obtain the full-length coding sequence (CDS) and 5′-flanking sequence of CYP19 gene, to investigate its expression and distribution profiles in different sized follicles, and to analyze the transcriptional regulatory mechanism of CYP19 gene in goose. Results showed that its CDS consisted of 1512 nucleotides and the encoded amino acid sequence contained a classical P450 structural domain. Homology analysis showed that there were high homologies of nucleotide and amino acid sequences between goose and other avian species. Its promoter sequence spanned from −1925 bp to the transcription start site (ATG) and several transcriptional factors were predicted in this region. Further analysis from luciferase assay showed that the luciferase activity was the highest spanning from −118 to −1 bp by constructing deletion promoter reporter vector. In addition, result from quantitative real-time polymerase chain reaction indicated that the mRNA level of CYP19 gene were highly expressed in theca layer of the fifth largest follicle, and the cellular location was in the theca externa cells by immunohistochemistry. Taken together, it could be concluded that the transcription activity of CYP19 gene was activated by transcriptional factors in its proximal region of promoter to promote the synthesis of estrogens, regulating the selection of pre-hierarchical into hierarchical follicle in goose.



中文翻译:

鹅卵泡中CYP19基因的分子特征、表达谱和转录调控

细胞色素 P450 家族 19 (CYP19) 是催化雄激素转化为雌激素的关键酶。然而,鹅CYP19基因的调控机制仍知之甚少。本研究试图获得CYP19基因的全长编码序列(CDS)和5'-侧翼序列,研究其在不同大小卵泡中的表达和分布特征,分析CYP19基因在鹅中的转录调控机制。结果表明,其CDS由1512个核苷酸组成,编码的氨基酸序列包含一个经典的P450结构域。同源性分析表明,鹅与其他鸟类的核苷酸和氨基酸序列具有较高的同源性。它的启动子序列从 -1925 bp 到转录起始位点 (ATG),并且在该区域预测了几种转录因子。荧光素酶测定进一步分析表明,通过构建缺失启动子报告载体,荧光素酶活性在-118至-1 bp范围内最高。此外,实时定量聚合酶链反应的结果表明CYP19基因的mRNA水平在第五大卵泡的膜层高表达,免疫组化显示细胞定位在外膜细胞。综上所述,CYP19基因的转录活性受其启动子近端区域的转录因子激活,促进雌激素的合成,

更新日期:2021-09-04
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