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Issue:ISSN 1000-7083
          CN 51-1193/Q
Director:Sichuan Association for Science and Technology
Sponsored by:Sichuan Society of Zoologists; Chengdu Giant Panda Breeding Research Foundation; Sichuan Association of Wildlife Conservation; Sichuan University
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Molecular cloning and expression analysis of cytochrome P450 (CYP370C2) gene in the white shrimp Litopenaeus vannamei
Author of the article:ZHENG PH, WANG L, ZHANG XX, WANG DM, LI JT, et al.
Author's Workplace:1. Hainan Provincial Key Laboratory for Functional Components Research and Utilization of Marine Bio-resources, Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan 571101, China; 2. Institute for Brain Research and Rehabilitation, South China Normal University, Guangzhou, Guangdong 510631, China 3. College of Life Science, South China Normal University, Guangzhou, Guangdong 510631, China
Key Words:cytochrome P450; Litopenaeus vannamei; ammonia; lipopolysaccharide
Abstract:Cytochrome P450 (CYP) enzyme system plays vital roles in metabolism of toxin, contaminant and medicine. In order to analyze the structure and functions of shrimp CYP, the full-length cDNA sequence of a CYP gene (named CYP370C2) was cloned from the Pacific white shrimp, Litopenaeus vannamei by using RACE technique. The full length cDNA of CYP370C2 was 1 754 bp, containing a 1 464 bp open reading frame which encoded 487 amino acids. The sequence of CYP370C2 shares common characteristics with CYP superfamily, such as heme-binging domain, I helix, C helix and K helix. The result of sequence alignment showed that the amino acid sequence of CYP370C2 shares 64% identity with that of the blue crab Portunus trituberculatus. Phylogenetic analysis showed that the CYP370C2 from L. vannamei was closely related to those from Malacostraca and Crustacea, suggesting that CYP gene is relatively conservative in aquatic arthropod. CYP370C2 were detected in all the detected tissues, and the highest expression was found in hepatopancrea, followed by gill and intestine. In the 48 h ammonia-N exposure experiment, the expression levels of CYP370C2 in both hepatopancrea and gill were significantly up-regulated after 24 h exposure, and the peak value appeared after 48 h and 24 h exposure respectively. After LPS injection, CYP370C2 transcription in hepatopancrea obviously increased after 3 h, and the highest level occurred after 12 h. Expression of CYP370C2 in gill was restrained after 3 and 6 h LPS injection, and then recovered after 12 h and up-regulated after 24 and 48 h. These results demonstrate that CYP370C2 participate in responses against ammonia-N stress and LPS stimulation, suggesting that CYP370C2 may play vital roles in L. vannamei defense system against environmental stress and pathogen infection.
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