title: Molecular cloning, expression and antioxidant characterisation of a typical thioredoxin gene (AccTrx2) in Apis cerana cerana.
abstract: Thioredoxins (Trxs) are a family of small, highly conserved and ubiquitous proteins that are involved in protecting organisms against toxic reactive oxygen species (ROS). In this study, a typical thioredoxin 2 gene was isolated from Apis cerana cerana, AccTrx2. The full-length cDNA sequence of AccTrx2 was composed of 407 bp containing a 318 bp open reading frame (ORF) that encodes a predicted protein of 105 amino acids, 11.974 kDa and an isoelectric point of 4.45. Expression profile of AccTrx2 as determined by a quantitative real-time PCR (qRT-PCR) analysis was higher in brain than in other tissues, with its highest transcript occurring on the 15day post-emergence adult and upregulated by such abiotic stresses as 4 °C, 16 °C, 25 °C, H2O2, cyhalothrin, acaricide, paraquat, phoxime and mercury (HgCl2) treatments. However, AccTrx2 was slightly repressed when exposed to 42 °C treatment. Characterisation of the recombinant protein showed that the purified AccTrx2 had insulin disulfide reductase activity and could protect DNA from ROS damage. These results indicate that AccTrx2 functions as an antioxidant that plays an important role in response to oxidative stress.
authors.affiliation: State Key Laboratory of Crop Biology, College of Life Sciences, Shandong Agricultural University, Taian, Shandong 271018, PR China.
authors.forename: Pengbo
authors.initials: P
authors.lastname: Yao
authors.forename: Lili
authors.initials: L
authors.lastname: Hao
authors.forename: Fang
authors.initials: F
authors.lastname: Wang
authors.forename: Xiaobo
authors.initials: X
authors.lastname: Chen
authors.forename: Yan
authors.initials: Y
authors.lastname: Yan
authors.forename: Xingqi
authors.initials: X
authors.lastname: Guo
authors.forename: Baohua
authors.initials: B
authors.lastname: Xu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001516:Bees / Q000235:genetics* / Q000378:metabolism; D001921:Brain / Q000378:metabolism; D003001:Cloning, Molecular; D004249:DNA Damage; D005786:Gene Expression Regulation; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010802:Phylogeny; D010957:Plasmids / Q000235:genetics; D017382:Reactive Oxygen Species / Q000378:metabolism; D013312:Stress, Physiological; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D019476:Insect Proteins; D017382:Reactive Oxygen Species; D013879:Thioredoxins; D010088:Oxidoreductases; C066352:coenzyme M-7-merceptoheptanoylthreonine-phosphate-heterodisulfide hydrogenase
keywords: 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; AccTrx2; Antioxidant capacity; Apis cerana cerana; DTT; EDTA; HEPES; IPTG; MFO; NTA; ORF; Oxidative damage; RACE-PCR; ROS; SDS-PAGE; Trxs; UTR; dithiothreitol; ethylenediaminetetraacetic acid; isopropyl-1-thio-β-galactopyranoside; mixed-function oxidation; nitrilotriacetic acid; open reading frame; qRT-PCR; quantitative real-time PCR; rapid amplification of cDNA ends PCR; reactive oxygen species; sodium dodecyl sulfate-polyacrylamide gel electrophoresis; thioredoxins; untranslated region