Verification and tissue-specific expression of nifU-like gene from the amphioxus Branchiostoma belcheri.

Lei Li, Shicui Zhang, Chunxin Fan, Zhenhui Liu, Jing Luan
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Abstract

A nifU-like gene exhibiting similarity to nifU of nitrogen fixation gene cluster was identified for the first time from the gut cDNA library of amphioxus Branchiostoma belcheri. Both RT-PCR and Northern blotting as well as in situ hybridization histochemistry verified that the cDNA represents an amphioxus nifU-like gene rather than a microbial contaminant. The nifU-like gene encodes a protein of 164 amino acid residues including a highly-conserved U-type motif (C-X(26)-C-X(43)-C), and shares 66-86% identity to NifU-like proteins from a variety of species including vertebrates, invertebrates and microbes. It is expressed in a tissue-specific manner in the digestive system including epipharyngeal groove, endostyle, hepatic caecum and hindgut and in the gill, ovary and testis. Taken together, it is highly likely that NifU-like protein plays some tissue-dependent and critical role in amphioxus.

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文昌鱼belcheri Branchiostoma nifU-like基因的验证及组织特异性表达。
首次从文昌鱼(amphioxus belcheri Branchiostoma)肠道cDNA文库中鉴定出一个与固氮基因簇nifU相似的nifU-like基因。RT-PCR和Northern blotting以及原位杂交组织化学证实,该cDNA代表文昌鱼nifu样基因,而不是微生物污染物。nifU-like基因编码一个包含164个氨基酸残基的蛋白,包括一个高度保守的u型基序(C-X(26)-C- x (43)-C),与来自脊椎动物、无脊椎动物和微生物等多种物种的nifU-like蛋白具有66-86%的同源性。它以组织特异性的方式表达于消化系统,包括咽沟、内腔、肝盲肠和后肠,以及鳃、卵巢和睾丸。综上所述,极有可能nifu样蛋白在文文鱼中起着组织依赖性的关键作用。
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