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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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Your Position :Home->Past Journals Catalog->2018 Vol.37 No.6

Molecular Cloning and Expression Analysis of Cytochrome P450 (CYP370C2) Gene in Litopenaeus vannamei
Author of the article:ZHENG Peihua1,3, WANG Lei2, ZHANG Xiuxia1, WANG Dongmei1, LI Juntao1, LU Yaopeng1,3, XIAN Jian'an1*, WANG Anli3*
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 571101, China;
2. Institute for Brain Research and Rehabilitation, South China Normal University, Guangzhou 510631, China;
3. College of Life Sciences, South China Normal University, Guangzhou 510631, China
Key Words:cytochrome P450; Litopenaeus vannamei; ammonia-N; lipopolysaccharide
Abstract:Cytochrome P450 (CYP) enzyme system plays vital roles in the metabolic processes 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 rapid-amplification of cDNA ends technique. The cDNA of CYP370C2 was 1 745 bp in length, containing a 1 464 bp open reading frame encoding 487 amino acids. The sequence of CYP370C2 shared common characteristics with the CYP superfamily, such as the heme-binging domain, Ⅰ helix, C helix and K helix. The result of sequence alignment showed that the amino acid sequence of CYP370C2 shared 64% identity with that of the blue crab (Portunus trituberculatus). Phylogenetic analysis showed that the CYP370C2 gene of L. vannamei was closely related to those of Malacostraca and Crustacea, indicating that CYP gene is relatively conserved in aquatic arthropod. The expression of CYP370C2 gene was detected in all the tissues of L. vannamei tested in this study, and the highest expression level was found in hepatopancrea, followed by gill and intestine. In the 48-h ammonia-N exposure experiment, the expression levels of CYP370C2 in hepatopancrea and gill were significantly upregulated after 24 h, and the peak value appeared after 48 h and 24 h, respectively. After lipopolysaccharide (LPS) injection, CYP370C2 transcription in hepatopancrea obviously increased after 3 h, and the highest level occurred after 12 h. By contrast, the expression of CYP370C2 in gill was inhibited after 3 h and 6 h, and then recovered after 12 h and upregulated after 24 h and 48 h. These results demonstrate that CYP370C2 participate in the host response against ammonia-N stress and LPS stimulation, and thus CYP370C2 may play vital roles in L. vannamei defense system against environmental stress and pathogen infection.
2018,37(6): 628-636 收稿日期:2018-04-25
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