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ZIRC
ZFIN ID: ZDB-PUB-080331-3
Identification and characterization of a novel gene differentially expressed in zebrafish cross-subfamily cloned embryos
Pei, D.S., Sun, Y.H., Chen, C.H., Chen, S.P., Wang, Y.P., Hu, W., and Zhu, Z.Y.
Date: 2008
Source: BMC Developmental Biology   8: 29 (Journal)
Registered Authors: Hu, Wei, Pei, Desheng, Sun, Yonghua, Zhu, Zuoyan
Keywords: none
MeSH Terms:
  • Animals
  • Cellular Reprogramming
  • Cloning, Organism/methods*
  • Crosses, Genetic
  • Embryo, Nonmammalian
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • In Situ Hybridization
  • Nuclear Transfer Techniques*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Up-Regulation
  • Zebrafish/embryology
  • Zebrafish/genetics*
  • Zebrafish Proteins/genetics*
PubMed: 18366661 Full text @ BMC Dev. Biol.
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ABSTRACT
BACKGROUND: Cross-species nuclear transfer has been shown to be a potent approach to retain the genetic viability of a certain species near extinction. However, most embryos produced by cross-species nuclear transfer were compromised because that they were unable to develop to later stages. Gene expression analysis of cross-species cloned embryos will yield new insights into the regulatory mechanisms involved in cross-species nuclear transfer and embryonic development. RESULTS: A novel gene, K31, was identified as an up-regulated gene in fish cross-subfamily cloned embryos using SSH approach and RACE method. K31 complete cDNA sequence is 1106 base pairs (bp) in length, wir transfer and embryonic development. RESULTS: A novel gene, K31, was identified as an th a 342 bp open reading frame (ORF) encoding a putative protein of 113 amino acids (aa). Comparative analysis revealed no homologous known gene in zebrafish and other species databacomplete cDNA sequence is 1106 base pairs (bp) in length, with a 342 bp open reading frame (ORF) encoding a putative protein of 113 amino ase. K31 protein contains a putative transmembrane helix and five putative phosphorylation sites but without a signal peptide. Expression pattern analysis by real time RT-PCR and whole-mount in situ hybridization (WISH) shows that it has the characteristics of constitutively expressed gene. Sub-cellular localization assay shows that K31 protein can not penetrate the nuclei. Interestingly, over-expression of K31 gene can cause lethality in the epithelioma papulosum cyprinid (EPC) cells in cell culture, which gave hint to the inefficient reprogramming events occurred in cloned embryos. CONCLUSIONS: Taken together, our findings indicated that K31 gene is a novel gene differentially expressed in fish cross-subfamily cloned embryos and over-expression of K31 gene can cause lethality of cultured fish cells. To our knowledge, this is the first report on the determination of novel genes involved in nucleo-cytoplasmic interaction of fish cross-subfamily cloned embryos.
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