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Gibel carp germ cell marker Vasa: cDNA cloning and its antibody preparation
Xu Hong-Yan (dbshyh@nus.edu.sg); Peng Jin-Xia; Gui Jian-Fang; Hong Yun-Han; Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol and Biotechnol, Wuhan 430072, Peoples R China
2005
Source PublicationActa Zoologica Sinica
ISSN0001-7302
Volume51Issue:4Pages:732-742
AbstractThe basis for animal reproduction and reproductive biotechnology is germ cells that are segregated from the somatic lineage early in embryonic development and produce sperm and eggs for germline transmission between generations. To provide a germ cell marker, we cloned and characterized Cagvasa, a vasa homolog from the gibel carp Carassius auratus gibelio. The Cagvasa cDNA is 2771 nt (nucleotide) and encodes a 701-aa (amino acid) protein (Cag Vasa) that possesses 14 RGG-repeats and all the eight conserved motifs of Vasa proteins. Cag Vasa shows sequence identities of 95%, 89%, 61%-66% and 50% to Vasa proteins from the common carp, zebrafish, tetrapods and Drosophila, respectively. RNA in situ hybridization on ovarian sections showed that Cagvasa was absent in any gonadal somatic cells but dynamically expressed throughout oogenesis: the signal is weak in oogonia, peaks in vitellogenic oocytes and deceases in maturing oocytes. To develop a tool for analyzing fish germ cells, we generated alpha Vasa, a rabbit antibody against a recombinant protein of 310-aa N-terminal Cag Vasa. Western blot analyses showed that alpha Vasa detected a 75 kD protein exclusively in the gibel carp gonads and isolated oocytes. To confirm the germ cell-specifity of alpha Vasa, fluorescent immunostaining was performed on gonadal sections. This antibody stains exclusively germ cells throughout oogenesis and early stages of spermatogenesis. We conclude that vasa encodes a Vasa ortholog that services a first germ cell marker in the gibel carp, and that aVasa provides a highly sensitive and powerful tool for the analysis of germ cell specification and differentiation in the gibel carp.
Indexed By其他
Document Type期刊论文
Identifierhttp://ir.ihb.ac.cn/handle/342005/18489
Collection期刊论文
Corresponding AuthorHong Yun-Han; Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol and Biotechnol, Wuhan 430072, Peoples R China
Recommended Citation
GB/T 7714
Xu Hong-Yan ,Peng Jin-Xia,Gui Jian-Fang,et al. Gibel carp germ cell marker Vasa: cDNA cloning and its antibody preparation[J]. Acta Zoologica Sinica,2005,51(4):732-742.
APA Xu Hong-Yan ,Peng Jin-Xia,Gui Jian-Fang,Hong Yun-Han,&Chinese Acad Sci, Inst Hydrobiol, State Key Lab Freshwater Ecol and Biotechnol, Wuhan 430072, Peoples R China.(2005).Gibel carp germ cell marker Vasa: cDNA cloning and its antibody preparation.Acta Zoologica Sinica,51(4),732-742.
MLA Xu Hong-Yan ,et al."Gibel carp germ cell marker Vasa: cDNA cloning and its antibody preparation".Acta Zoologica Sinica 51.4(2005):732-742.
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