title: Full-length cDNA cloning, molecular characterization and differential expression analysis of peroxiredoxin 6 from Ovis aries.
abstract: Peroxiredoxin 6 (Prdx6), an important antioxidant enzyme that can eliminate reactive oxygen species (ROS) to maintain homeostasis, is a bifunctional protein that possesses the activities of both glutathione peroxidase and phospholipase A2. In this study, a novel full-length Prdx6 cDNA (OaPrdx6) was cloned from Sheep (Ovis aries) using rapid amplification of cDNA ends (RACE). The full-length cDNA of OaPrdx6 was 1753bp containing a 5'-untranslated region (UTR) of 93bp, a 3'-UTR of 985bp with a poly(A) tail, and an open reading frame (ORF) of 675bp encoding a protein of 224 amino acid residues with a predicted molecular weight of 25.07kDa. The recombinant protein OaPrdx6 was expressed and purified, and its DNA protection activity was identified. In order to analyze the Prdx6 protein expression in tissues from O. aries, monoclonal antibodies against OaPrdx6 were prepared. Western blotting results indicated that OaPrdx6 protein could be detected in heart, liver, spleen, lung, kidney, stomach, intestine, muscle, lymph node and white blood cells, and the highest expression was found in lung while the lowest expression in muscle. Compared to the normal sheep group, the mRNA transcription level of Prdx6 in buffy coat was up-regulated in the group infected with a virulent field strain of Brucella melitensis, and down-regulated in the group inoculated with a vaccine strain S2 of brucellosis. The results indicated that Prdx6 was likely to be involved in the host immune responses against Brucella infection, and probably regarded as a molecular biomarker for distinguishing between animals infected with virulent Brucella infection and those inoculated with vaccine against brucellosis.
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education/Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
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, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
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, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
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, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
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, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Yan-Song
authors.initials: YS
authors.lastname: Li
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education/Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Xiao-Li
authors.initials: XL
authors.lastname: Feng
authors.affiliation: Jilin Provincial Center for Animal Disease Control and Prevention, Changchun 130062, China.
authors.forename: Shou-Yin
authors.initials: SY
authors.lastname: Zhang
authors.affiliation: Jilin Provincial Center for Animal Disease Control and Prevention, Changchun 130062, China.
authors.forename: Nan
authors.initials: N
authors.lastname: Wang
authors.affiliation: Jilin Entry-Exit Inspection and Quarantine Bureau, Changchun 130062, China.
authors.forename: Qing-Feng
authors.initials: QF
authors.lastname: Meng
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education/Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Yong-Jie
authors.initials: YJ
authors.lastname: Yang
authors.affiliation: College of Animal Husbandry and Veterinary, Liaoning Medical University, Jinzhou 121001, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Tang
authors.affiliation: Department of Husbandry and Veterinary Medicine, Jiangsu Polytechnic College of Agriculture and Forestry, Jurong 212400, China.
authors.forename: Yun-Ming
authors.initials: YM
authors.lastname: Xu
authors.affiliation: Fujian Institute of Subtropical Botany, Xiamen 361006, China.
authors.forename: Wen-Hui
authors.initials: WH
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education/Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Xing
authors.initials: X
authors.lastname: Guo
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education/Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
authors.forename: Xiao-Feng
authors.initials: XF
authors.lastname: Chen
authors.affiliation: Key Laboratory of Zoonosis Research, Ministry of Education/Institute of Zoonosis/College of Veterinary Medicine, Jilin University, Xi An Da Lu 5333, Changchun 130062, China.
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, Jilin University, Xi An Da Lu 5333, Changchun 130062, China. Electronic address: renhl@jlu.edu.cn.
authors.forename: Hong-Lin
authors.initials: HL
authors.lastname: Ren
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D001483:Base Sequence; D017347:Brucella melitensis / Q000276:immunology; D002006:Brucellosis / Q000276:immunology / Q000662:veterinary; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D054465:Peroxiredoxin VI / Q000235:genetics* / Q000276:immunology; D011994:Recombinant Proteins / Q000276:immunology; D012756:Sheep
chemical_list: D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D011994:Recombinant Proteins; D054465:Peroxiredoxin VI
keywords: Brucellosis; Expression analysis; Monoclonal antibodies; Ovis aries; Peroxiredoxin 6