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Cloning, Tissue-specific Expression and Evolutional Analysis of CNBP and eIF-5A in Grass Carp

Author: ChenQiong
Tutor: LinGang
School: Nanchang University
Course: Zoology
Keywords: Grass carp Cell nucleic acid binding protein ( CNBP ) Eukaryotic translation initiation factor 5A (eIF-5A) Expression Evolution
CLC: Q78
Type: Master's thesis
Year: 2008
Downloads: 21
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Abstract


From grass carp (Ctenopharyngodon idella) liver and kidney cDNA library and cloned into the grass carp cell nucleic acid binding protein and eukaryotic translation initiation 5A . Grass carp cell nucleic acid binding protein (Cellular nucleic acid-binding protein, CNBP) full-length cDNA is 1948bp . Including the 5 ' untranslated region of 33 bp in the 3' untranslated region of 1423 bp . The open reading frame of 492 bp, encoding 163 amino acids . The grass carp CNBP contains seven conserved CCHC -type zinc finger structure , nuclear localization signal and RGG box high homology with other fish . And other vertebrates , compared to of grass carp CNBP sites in the 25 amino acid substitution , wherein the first five amino acid residues in the three zinc finger by Gly becomes conserved exist in fish HIS Further , deletions of 6 to 14 amino acid residues in zinc finger 1 , and the second zinc finger structures . However , the difference of this structure is not enough to produce the new features , because adaptive evolution analysis showed that cell nucleic acid binding protein did not experience positive Darwinian selection ( ω ≤ 1 ) . RT-PCR was experimental results show that , in the grass carp heart in normal grass carp liver , intestine , kidney , spleen , heart , muscle , gills and brain eight kinds of organizations CNBP expression more significant , followed by brain , gill , muscle , in the kidney also weak expression . Grass carp eukaryotic translation initiation factor 5A ( eukaryotic translation initiations for factor 5A , eIF -5A ) full-length cDNA of 2249bp, which contains 5 'untranslated region of 41bp 3' non - coding region of 1743bp, encoding eIF-5A 155 amino acid residues molecular weight 16.856ku , and pI 5.18 . The N-terminus of the grass carp eIF-5A protein containing eIF-5A family the carboxylation putrescine Total lysine in the protein C-terminus contains a leucine - rich domain . Grass carp eIF-5A having a high homology with human and other animals eIF-5A , and only a small number of amino acid residue substitution . Adaptive evolution analysis also shows that not only has the conservative nature of the structure of eIF-5A also has the function of a conservative . eIF-5A constant expression the grass carp heart , brain , muscle , gills , spleen , liver , intestine , kidney and other tissues , and Poly I: C induced expression does not change .

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