cDNA cloning, characterization and expression analysis of catalase in swimming crab Portunus trituberculatus: cDNA cloning and expression analysis of catalase gene of Portunus trituberculatus.
Catalase is an important antioxidant protein that protects organisms against various oxidative stresses by eliminating hydrogen peroxide. In the present study, a full-length cDNA sequence of catalase was cloned from the haemocytes of swimming crab Portunus trituberculatus by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end method. The catalase cDNA sequence contained 1,851 bp with an open reading frame of 1,551 bp encoding 516 amino acid residues. The conserved catalytic active residues His-71, Asn-144 and Tyr-354 were predicted in the amino acid sequence of P. trituberculatus catalase. The deduced catalase protein had a calculated molecular mass of 58.5 kDa with an estimated isoelectric point of 6.90. Multiple alignment analysis revealed that the deduced amino acid sequence of catalase shared high identity of 68-95 % with those of other species. Quantitative real-time RT-PCR analysis revealed that P. trituberculatus catalase transcript was strongly detected in haemocytes, hepatopancreas, heart, stomach, intestine, gill, ovary and muscle. The expression level of catalase transcripts both in haemocytes and hepatopancreas changed rapidly and dynamically after Vibrio alginolyticus challenging. These facts indicate that catalase was perhaps involved in the acute response against invading bacteria and was an inducible protein involved in the host innate immune response through elimination of H(2)O(2) in crab.
Key Laboratory for Sustainable Utilization of Marine Fisheries Resources, Ministry of Agriculture, 106 Nanjing Road, Qingdao 266071, People's Republic of China.
Ping
P
Chen
Jitao
J
Li
Ping
P
Liu
Baoquan
B
Gao
Qingyin
Q
Wang
Jian
J
Li
D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003386:Brachyura / Q000201:enzymology* / Q000382:microbiology; D002374:Catalase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D015870:Gene Expression; D015971:Gene Expression Regulation, Enzymologic; D006434:Hemocytes / Q000201:enzymology; D043143:Hepatopancreas / Q000201:enzymology; D054884:Host-Pathogen Interactions; D007113:Immunity, Innate; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D044149:Vibrio alginolyticus / Q000502:physiology
D060829:Arthropod Proteins; D018076:DNA, Complementary; D002374:Catalase