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Molecular cloning and expression of kidney-type glutaminase from common carp (Cyprinus carpio) and its up-regulation by glutamine in primary culture enterocyte

Authors

  • J. Jiang,

    1. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, China
    2. Fish Nutrition and Safety Production, University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China
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  • L. Feng,

    1. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, China
    2. Fish Nutrition and Safety Production, University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China
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  • Y. Liu,

    1. Fish Nutrition and Safety Production, University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China
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  • W.-D. Jiang,

    1. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, China
    2. Fish Nutrition and Safety Production, University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China
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  • K. Hu,

    1. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, China
    2. Fish Nutrition and Safety Production, University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China
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  • S.-H. Li,

    1. Fish Nutrition and Safety Production, University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China
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  • X.-Q. Zhou

    Corresponding author
    1. Animal Nutrition Institute, Sichuan Agricultural University, Chengdu, China
    2. Fish Nutrition and Safety Production, University Key Laboratory of Sichuan Province, Sichuan Agricultural University, Chengdu, China
    3. Key Laboratory for Animal Disease-Resistance Nutrition of China Ministry of Education, Sichuan Agricultural University, Chengdu, China
    • Correspondence: X.-Q. Zhou, Animal Nutrition Institute, Sichuan Agricultural University, Chengdu 611130, China. E-mail: zhouxq@sicau.edu.cn

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Abstract

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney-type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real-time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up-regulated quickly and effectively by Gln.

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