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Int. J. Mol. Sci. 2016, 17(8), 1206; doi:10.3390/ijms17081206

Full-Length cDNA Cloning, Molecular Characterization and Differential Expression Analysis of Lysophospholipase I from Ovis aries

1
Key Laboratory of Zoonosis Research, Ministry of Education/Institute of Zoonosis/College of Veterinary Medicine, College of Animal Sciences, Jilin University, Xi An Da Lu 5333, Changchun 130062, China
2
Department of Food Science, College of Agriculture, Yanbian University, Yanji 133002, China
3
Animal Husbandry and Veterinary Unit of Xiangyang Town, Liuhe 135305, China
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editors: Mateus Webba da Silva and Ritva Tikkanen
Received: 3 May 2016 / Revised: 15 June 2016 / Accepted: 19 July 2016 / Published: 28 July 2016
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

Lysophospholipase I (LYPLA1) is an important protein with multiple functions. In this study, the full-length cDNA of the LYPLA1 gene from Ovis aries (OaLypla1) was cloned using primers and rapid amplification of cDNA ends (RACE) technology. The full-length OaLypla1 was 2457 bp with a 5′-untranslated region (UTR) of 24 bp, a 3′-UTR of 1740 bp with a poly (A) tail, and an open reading frame (ORF) of 693 bp encoding a protein of 230 amino acid residues with a predicted molecular weight of 24,625.78 Da. Phylogenetic analysis showed that the OaLypla1 protein shared a high amino acid identity with LYPLA1 of Bos taurus. The recombinant OaLypla1 protein was expressed and purified, and its phospholipase activity was identified. Monoclonal antibodies (mAb) against OaLypla1 that bound native OaLypla1 were generated. Real-time PCR analysis revealed that OaLypla1 was constitutively expressed in the liver, spleen, lung, kidney, and white blood cells of sheep, with the highest level in the kidney. Additionally, the mRNA levels of OaLypla1 in the buffy coats of sheep challenged with virulent or avirulent Brucella strains were down-regulated compared to untreated sheep. The results suggest that OaLypla1 may have an important physiological role in the host response to bacteria. The function of OaLypla1 in the host response to bacterial infection requires further study in the future. View Full-Text
Keywords: Lysophospholipase I; Ovis aries; Brucella; tissue distribution; differential expression Lysophospholipase I; Ovis aries; Brucella; tissue distribution; differential expression
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Liu, N.-N.; Liu, Z.-S.; Hu, P.; Zhang, Y.; Lu, S.-Y.; Li, Y.-S.; Yang, Y.-J.; Zhang, D.-S.; Zhou, Y.; Ren, H.-L. Full-Length cDNA Cloning, Molecular Characterization and Differential Expression Analysis of Lysophospholipase I from Ovis aries. Int. J. Mol. Sci. 2016, 17, 1206.

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