Molecular cloning and expression of NOS in shrimp, Litopenaeus vannamei

被引:61
作者
Yao, Cui-Luan [2 ]
Ji, Pei-Feng [2 ]
Wang, Zhi-Yong [2 ]
Li, Fu-hua [1 ]
Xiang, Jian-Hai [1 ]
机构
[1] Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
[2] Jimei Univ, Fisheries Biotechnol Inst, Coll Fisheries, Fujian Prov Univ,Key Lab Sci & Technol Aquacultur, Xiamen 361021, Peoples R China
基金
中国国家自然科学基金;
关键词
NOS; Litopenaeus vannamei; Expression; LPS; Poly I C; NITRIC-OXIDE SYNTHASE; FENNEROPENAEUS-CHINENSIS; LAMINARIN STIMULATION; ARGININE KINASE; GENE-EXPRESSION; CHINESE SHRIMP; MESSENGER-RNA; INOS GENE; HEMOLYMPH; RAT;
D O I
10.1016/j.fsi.2009.12.002
中图分类号
S9 [水产、渔业];
学科分类号
0908 ;
摘要
The importance of the nitric oxide synthase (NOS) gene family is demonstrated by many studies in recent years However, the lack of sequence information and clones of shrimp NOS cDNA limits further study on its characterization and function in this species In this report, the cDNA of NOS contained full-length ORF was cloned from the Pacific white shrimp, Litopenaeus vannamei It was of 4680 bp, including a 5'-terminal untranslated region (UTR) of 278 bp, a 3'-terminal UTR of 862 bp, which contained 5 ATTTA repeats, and an open reading frame (ORF) of 3540 bp encoding a polypeptide of 1179 amino acids It contained a typical NO synthase domain at the N-terminal, next to a flavodoxin I domain, a flavin adenine dinucleotide (FAD) binding domain, respectively, and a conservative nicotinamide adenine dinucleotide (NAD) binding domain structure at the C-terminal Quantitative real-time reverse transcription PCR analysis revealed L vannamei NOS (LvNOS) to be expressed in most shrimp tissues, with highest expression in the hepatopancreas and weakest expression in skin. The expression of LvNOS after challenge with LPS and poly I:C was tested in hemocytes, hepatopancreas and nerve The results indicated that the NOS transcript level could be induced in hemocytes by injection with LPS The highest expression was in the hemocyte. with 8 8 times (at 3 h) as much as that in the control (p < 0.05) However, sharp clown-regulation of NOS was found in hepatopancreas and nerve after LPS and poly I-C injection (p < 0 05). These results Suggested that NOS might play an important role in shrimp's defense against pathogenic infection. (C) 2009 Elsevier Ltd All rights reserved
引用
收藏
页码:453 / 460
页数:8
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