title: Full-Length cDNA Cloning, Molecular Characterization and Differential Expression Analysis of Lysophospholipase I from Ovis aries.
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.
authors.affiliation: 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. liunannan112117@163.com.
authors.forename: Nan-Nan
authors.initials: NN
authors.lastname: Liu
authors.affiliation: 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. zsliu1959@163.com.
authors.forename: Zeng-Shan
authors.initials: ZS
authors.lastname: Liu
authors.affiliation: 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. hupan84@163.com.
authors.forename: Pan
authors.initials: P
authors.lastname: Hu
authors.affiliation: 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. zhangying201604@163.com.
authors.forename: Ying
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: 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. lushiying1129@163.com.
authors.forename: Shi-Ying
authors.initials: SY
authors.lastname: Lu
authors.affiliation: 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. l_ys92305@163.com.
authors.forename: Yan-Song
authors.initials: YS
authors.lastname: Li
authors.affiliation: Department of Food Science, College of Agriculture, Yanbian University, Yanji 133002, China. yjyang@ybu.edu.cn.
authors.forename: Yong-Jie
authors.initials: YJ
authors.lastname: Yang
authors.affiliation: Animal Husbandry and Veterinary Unit of Xiangyang Town, Liuhe 135305, China. zhangdongsong2016@163.com.
authors.forename: Dong-Song
authors.initials: DS
authors.lastname: Zhang
authors.affiliation: 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. zhouyu69@sina.com.
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.affiliation: 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. renhl@jlu.edu.cn.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Ren
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D017347:Brucella melitensis / Q000201:enzymology / Q000235:genetics*; D002006:Brucellosis / Q000276:immunology / Q000662:veterinary; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D005260:Female; D008245:Lysophospholipase / Q000235:genetics*; D008807:Mice, Inbred BALB C; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D060888:Real-Time Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D012756:Sheep
chemical_list: D018076:DNA, Complementary; D008245:Lysophospholipase
keywords: Brucella; Lysophospholipase I; Ovis aries; differential expression; tissue distribution