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利用抑制性消减杂交技术鉴定克隆胚胎与合子发育的斑马鱼(Danio rerio)胚胎在穹顶期差异表达的基因。

Identification of differentially expressed genes between cloned and zygote-developing zebrafish (Danio rerio) embryos at the dome stage using suppression subtractive hybridization.

作者信息

Luo Daji, Hu Wei, Chen Shangping, Xiao Yi, Sun Yonghua, Zhu Zuoyan

机构信息

College of Life Sciences, Wuhan University, Wuhan, China.

出版信息

Biol Reprod. 2009 Apr;80(4):674-84. doi: 10.1095/biolreprod.108.074203. Epub 2008 Dec 17.

DOI:10.1095/biolreprod.108.074203
PMID:19092113
Abstract

Comparative analyses of differentially expressed genes between somatic cell nuclear transfer (SCNT) embryos and zygote-developing (ZD) embryos are important for understanding the molecular mechanism underlying the reprogramming processes. Herein, we used the suppression subtractive hybridization approach and from more than 2900 clones identified 96 differentially expressed genes between the SCNT and ZD embryos at the dome stage in zebrafish. We report the first database of differentially expressed genes in zebrafish SCNT embryos. Collectively, our findings demonstrate that zebrafish SCNT embryos undergo significant reprogramming processes during the dome stage. However, most differentially expressed genes are down-regulated in SCNT embryos, indicating failure of reprogramming. Based on Ensembl description and Gene Ontology Consortium annotation, the problems of reprogramming at the dome stage may occur during nuclear remodeling, translation initiation, and regulation of the cell cycle. The importance of regulation from recipient oocytes in cloning should not be underestimated in zebrafish.

摘要

比较体细胞核移植(SCNT)胚胎和受精卵发育(ZD)胚胎之间差异表达基因,对于理解重编程过程的分子机制至关重要。在此,我们采用抑制性消减杂交方法,从2900多个克隆中鉴定出斑马鱼穹顶期SCNT胚胎和ZD胚胎之间96个差异表达基因。我们报告了首个斑马鱼SCNT胚胎差异表达基因数据库。总体而言,我们的研究结果表明,斑马鱼SCNT胚胎在穹顶期经历了显著的重编程过程。然而,大多数差异表达基因在SCNT胚胎中表达下调,表明重编程失败。基于Ensembl描述和基因本体联合会注释,穹顶期重编程问题可能发生在核重塑、翻译起始和细胞周期调控过程中。在斑马鱼克隆中,受体卵母细胞调控的重要性不应被低估。

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