Molecular cloning, expression of a peroxiredoxin gene in Chinese shrimp Fenneropenaeus chinensis and the antioxidant activity of its recombinant protein.
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 942bp with a 594bp open reading frame, encoding 198 amino acids. The molecular mass of the deduced amino acid is 22041.17Da 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(2)O(2) in the presence of dithiothreitol.
Zhang
Qingli
Q
Key Laboratory of Experimental Marine Biology, Institute of Oceanology, Chinese Academy of Sciences, 7 Nanhai Road, Qingdao 266071, People's Republic of China.
Li
Fuhua
F
Zhang
Jiquan
J
Wang
Bing
B
Gao
Hongwei
H
Huang
Bingxin
B
Jiang
Hao
H
Xiang
Jianhai
J
D000595:Amino Acid Sequence; D000818:Animals; D000975:Antioxidants / Q000378:metabolism*; D001483:Base Sequence; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic; D006434:Hemocytes / Q000201:enzymology / Q000382:microbiology; D043143:Hepatopancreas / Q000201:enzymology / Q000382:microbiology; D006861:Hydrogen Peroxide / Q000378:metabolism; D008969:Molecular Sequence Data; D033561:Penaeidae / Q000201:enzymology* / Q000235:genetics* / Q000382:microbiology; D010455:Peptides / Q000737:chemistry; D010544:Peroxidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D054464:Peroxiredoxins; D010802:Phylogeny; D017510:Protein Folding; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D014733:Vibrio / Q000502:physiology; D014735:Vibrio Infections
D000975:Antioxidants; D010455:Peptides; D012333:RNA, Messenger; D011994:Recombinant Proteins; D006861:Hydrogen Peroxide; D010544:Peroxidases; D054464:Peroxiredoxins