Molecular cloning, expression of a peroxiredoxin gene in Chinese shrimp Fenneropenaeus chinensis and

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时间:2008-01-14  来源:文本大小:【 |  | 】  【打印
Qingli Zhang a,b, Fuhua Li a, Jiquan Zhang a, Bing Wang a, Hongwei Gaoa,b,
Bingxin Huanga,b, Hao Jiang a,b, Jianhai Xiang a,
a Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences,
7 Nanhai Road, Qingdao 266071, People’s Republic of China
b Graduate School, Chinese Academy of Sciences, Beijing 100039, People’s Republic of China
Received 23 January 2007; received in revised form 12 March 2007; accepted 14 March 2007
Available online 3 May 2007
Abstract
Peroxiredoxin (Prx) is known to be an antioxidant protein that protects the organisms against various oxidative stresses and functions inintracellular signal transduction. A Prx gene was firstly isolated in the crustacean, Chinese shrimp Fenneropenaeus chinensis. The full-lengthcDNA consists of 942 bp with a 594 bp open reading frame, encoding 198 amino acids. The molecular mass of the deduced amino acid is22041.17 Da with an estimated pI of 5.17. Sequence comparison showed that Prx of F. chinensis shares 76%, 73% and 72% identity with that ofAedes aegypti, Branchiostoma belcheri tsingtaunese and Drosophila melanogaster, respectively. Northern blot analysis revealed the presence ofPrx transcripts of F. chinensis in all tissues examined. Real-time PCR analysis indicated that the Prx showed different expression profiles in shrimphemocytes and hepatopancreas after artificial infection with Vibrio anguillarum. In addition, a fusion protein containing Prx was produced in vitro.LC–ESI–MS analysis showed that four peptide fragments of the recombinant protein were identical to the corresponding sequence of F. chinensisPrx. And the purified recombinant proteins were shown to reduce H2O2 in the presence of dithiothreitol.
© 2007 Elsevier Ltd. All rights reserved.
Keywords: Peroxiredoxin (Prx) gene; Expression profile; Vibrio anguillarum infection; Immune response; Recombinant protein; Activity; Fenneropenaeus chinensis
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