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KMID : 0381120150370100883
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Genes and Genomics
2015 Volume.37 No. 10 p.883 ~ p.892
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cDNA cloning, expression, and immunolocalization of gonad-inhibiting hormone (GIH) in Litopenaeus vannamei
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Li Guang-Li
Deng Si-Ping
Jiang Shu-Na
Ye Man
Chen Hua-Pu
Chan Siuming F.
Zhu Chun-Hua
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Abstract
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In this study, the full-length GIH cDNA sequence from Litopenaeus vannamei was cloned from the eyestalk by reverse transcriptase polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends. The full-length GIH cDNA was 865 bp with a 288 bp open-reading frame, which encoded a 96 amino acid prepro-GIH with 17 amino acid signal peptide. L. vannamei GIH (LvGIH) can be classified as a member of type-II crustacean hyperglycemic hormone polypeptide family. LvGIH shares 93.8 and 66.7 % amino acid sequence identity with GIH from Penaeus monodon, and the molt-inhibiting hormone from Marsupenaeus japonicas, respectively. By quantitative real-time PCR (qPCR), LvGIH mRNA transcripts were detected in fertilized eggs, nauplius, zoea, mysis and juveniles of 25, 35 and 40 days old. LvGIH transcript levels increased significantly with the development from fertilized eggs to juveniles. LvGIH transcript levels were highest in juveniles at 35 days old. By RT-PCR, LvGIH mRNA transcripts were detected only in the eyestalks and brains but not in the muscles, intestines, gills, heart, hepatopancreas, ovaries and testes of adults, and there was no difference in the expression level of LvGIH between males and females. Using the P. monodon anti-GIH antibody, we showed that LvGIH was located mainly in the XO-SG and slightly in axon, with similar fluorescence intensity found in XO and SG. To summarize, we have cloned and characterized the GIH of the shrimp L. vannamei. In addition to the GIH properties described in other crustaceans, a peak of LvGIH expression was identified at the time of sexual differentiation (i.e., day 35 larvae) suggesting that LvGIH may also be involved in the control of this process.
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KEYWORD
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Litopenaeus vannamei, Gonad-inhibiting hormone, Cloning, Expression
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