Molecular cloning, expression of a peroxiredoxin gene in Chinese shrimp Fenneropenaeus chinensis and the antioxidant activity of its recombinant protein
Autor(es): Zhang, Qingli; Li, Fuhua; Zhang, Jiquan; Wang, Bing; Gao, Hongwei; Huang, Bingxin; Jiang, Hao; Xiang, Jianhai
Resumo: Peroxiredoxin (Prx) is known to be an antioxidant protein that protects the organisms against various oxidative stresses and functions in intracellular signal transduction. A Prx gene was firstly isolated in the crustacean, Chinese shrimp Fenneropenaeus chinensis. The full-length cDNA consists of 942 bp with a 594 bp open reading frame, encoding 198 amino acids. The molecular mass of the deduced amino acid is 22041.17 Da with an estimated pI of 5.17. Sequence comparison showed that Prx of F. chinensis shares 76%, 73% and 72% identity with that of Aedes aegypti, Branchiostoma belcheri tsingtaunese and Drosophila melanogaster, respectively. Northern blot analysis revealed the presence of Prx transcripts of F. chinensis in all tissues examined. Real-time PCR analysis indicated that the Prx showed different expression profiles in shrimp hemocytes 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. chinensis Prx. And the purified recombinant proteins were shown to reduce H sub(2)O sub(2) in the presence of dithiothreitol.
Palavras-Chave: Gene expression; Recombinants; Antioxidants; Molecular weight; Proteins; Peroxiredoxin; Infection; Fenneropenaeus chinensis
Imprenta: Molecular Immunology, v. 44, n. 14, p. 3501-3509, 2007.
Descritores: Aedes aegypti - Immune response ; Aedes aegypti - Proteins ; Aedes aegypti - Real Time PCR ; Aedes aegypti - Immunology
Data de publicação: 2007
