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Molecular cloning and functional characterization of swine sodium dependent phosphate cotransporter type II b (NaPi-IIb) gene

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Abstract

A sodium-dependent phosphate transporter gene, NaPi-IIb, was isolated from swine small intestine using cDNA library screening method. Sequencing analysis revealed that the NaPi-IIb cDNA sequences was 2,016 bp in length and encoded an open-reading frame consisting of 671 amino acids. The cDNA showed 83.1 % sequences identity to the human NaPi-IIb and 78.7 % sequences identity to the chicken NaPi-IIb. Prediction of membrane spanning domains based on the hydrophilic and hydrophobic properties of the amino acids suggested that a putative protein had nine transmembrane domains, with both the NH2 and COOH terminal being intracellular. By northern blot, a ~4.2 kb transcript was found to be abundantly expressed in mall intestine, lung, ovary, mammary glands, liver, kidney, salivary glands, placenta and thymus. Microinjection of swine NaPi-IIb cRNA into Xenopus oocytes demonstrated that the NaPi-IIb showed sodium-dependent Pi cotransport activity, and an approximate 31-fold increase of Pi uptake was seen in cRNA injected oocytes. The swine NaPi-IIb transporter expressed in Xenopus oocytes had a Km for Pi of ~79.35 ± 7.2 μM. Furthermore, the pH dependency characterization of swine NaPi-IIb transporter showed activation at extracellular alkaline-pH.

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Acknowledgment

This research was supported by Grants from the National Natural Science Foundation of China (No.31172218), National Scientific and Technological Supporting Project (No. 2011BAD26B03), the Program for Science and Technology Innovative Research Team in Higher Educational Institutions of Hunan Province.

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Authors and Affiliations

  1. College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, Hunan Province, China

    Xiang Zhifeng, Fang Rejun, Hu Longchang & Su Wenqing

  2. College of Animal Science, Henan Institute of Science and Technology, Xinxiang, 453003, Henan Province, China

    Xiang Zhifeng

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  1. Xiang Zhifeng
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Correspondence to Fang Rejun.

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Zhifeng, X., Rejun, F., Longchang, H. et al. Molecular cloning and functional characterization of swine sodium dependent phosphate cotransporter type II b (NaPi-IIb) gene. Mol Biol Rep 39, 10557–10564 (2012). https://doi.org/10.1007/s11033-012-1941-0

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  • DOI: https://doi.org/10.1007/s11033-012-1941-0

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