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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
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Your Position :Home->Past Journals Catalog->2014 Vol.33 No.4

Cloning and Characterization of Inducible Nitric Oxide Synthase from Plateau Pika (Ochotona curzoniae)
Author of the article:XIE Ling1,2, GUO Xinyi1,2, ZHANG Xuze3, PANG Bo1,2, DU Yurong4, ZOU Xiaoyan1, GUO Songchang1*, ZHAO Xinquan1*
Author's Workplace:(1. Key Laboratory of Adaptation and Evolution of Plateau Biota, Northwest Institute of Plateau Biology, Chinese Academy of Sciences, Xining 810008, China; 2. University of Chinese Academy of Sciences, Beijing 100049, China; 3. Qinghai University for Nationalities, Xining 810007, China; 4. School of Life and Geography Science, Qinghai Normal University, Xining 810008, China)
Key Words:plateau pika; inducible nitric oxide synthase (iNOS); cDNA clone
Abstract:Plateau pika (Ochotona curzoniae), a native species living at 3000~5000 mabove sea-level on the Qinghai-Tibetan Plateau, is a typical hypoxia-tolerant mammal. Nitric oxide (NO), which has been proved to be a vasodilator, plays important roles in preventing hypoxia-induced hypoxic pulmonary vasoconstriction and pulmonary hypertension under hypoxia. Inducible nitric oxide synthase (iNOS) is a key enzymes that catalyzing L-arginine to NO and regulated by hypoxia stress. In this study, the cDNA of iNOS was cloned by RT-PCR and3’-rapid amplification PCR of cDNA ends (3’-RACE) from the lung of plateau pika. The results showed that the length of iNOS cDNA of plateau pika was 3981 bp, containing an open reading frame (ORF) of 3450 bp and encoding 1149 amino acid residues. The deduced amino acid sequence was highly identical to those of the American pika (98%), followed by rabbit (87%), human (82%), rat (78%), mouse (78%), dog (82%) and pig (83%). The typical iNOS domains, including oxidase domain, reductase domain and FAD binding site, were identified in the iNOS of plateau pika. The molecular phylogenetic trees based on ML and Bayesian methods showed that pika has a close relationship with rabbit. Three positive selection sites, 32T, 33Y and 46R, were identified by the PAML analysis. However, the modeling results showed that the evolution of iNOS was mainly under negative selection, suggested that there was no adaptive evolution in plateau pika iNOS. This study laid a foundation for further exploration on the expression pattern, function and regulatory mechanism of iNOS in plateau pika adapting to high altitude environment.
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