Molecular cloning, expression and antioxidant characterisation of a typical thioredoxin gene (AccTrx2) in Apis cerana cerana

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• 作者：Pengbo Yao ; Lili Hao ; Fang Wang ; Xiaobo Chen ; Yan Yan ; Xingqi Guo ; Baohua Xu
• 关键词：ROS ; reactive oxygen species ; Trxs ; thioredoxins ; UTR ; untranslated region ; ORF ; open reading frame ; RACE-PCR ; rapid amplification of cDNA ends PCR ; qRT-PCR ; quantitative real-time PCR ; IPTG ; isopropyl-1-thio-¦Â-galactopyranoside ; EDTA ; ethylenediaminetet
• 刊名：Gene
• 出版年：2013
• 出版时间：15 September, 2013
• 年：2013
• 卷：527
• 期：1
• 页码：33-41
• 全文大小：869 K

文摘

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 15 day post-emergence adult and upregulated by such abiotic stresses as 4 ¡ãC, 16 ¡ãC, 25 ¡ãC, H₂O₂, cyhalothrin, acaricide, paraquat, phoxime and mercury (HgCl₂) treatments. However, AccTrx2 was slightly repressed when exposed to 42 ¡ãC 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.