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Molecular cloning, characterization and expression analysis of thrombospondin gene from Penaeus monodon

机译:斑节对虾血小板反应蛋白基因的分子克隆,表征及表达分析

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In present study, a thrombospondin gene was obtained from the ovary and neurosecretory organ in eyestalk cDNA library of black tiger prawn (Penaeus monodon). The full-length P. monodon thrombospondin (PmTSP) cDNA contained a 5 untranslated region (UTR) of 9 bp, an open reading frame (ORF) of 2778 bp encoding a polypeptide of 925 amino acids with molecular mass 100.57 kDa, and a 3UTR of 99 bp. ScanProsite analysis indicated that PmTSP contained four chitin-binding type-II domains, an EGF-like domain, eight thrombospondin type-III repeats and one thrombospondin C-terminal domain. Homology analysis of the deduced amino acid sequence of the PmTSP with other known TSP sequences by MatGAT software revealed that the PmTSP shows very high homology with the sequences of Fennerpenaeus chinensis (89.9% similarity, 83.8% identity). Analysis of the tissue expression pattern of the PmTSP gene showed that the PmTSP mRNA was expressed in all tested tissues, including hepatopancreas, ovary, muscle, intestine, neurosecretory organ in eyestalk, neurosecretory organ in brain, stomach, and heart, with highest level in the ovary. Furthermore, the PmTSP expression was found to be of high level in six development stages of the ovary. The results indicated that PmTSP might play an important role in ovarian development.
机译:在本研究中,从黑虎虾(Penaeus monodon)的眼球cDNA文库中的卵巢和神经分泌器官获得了血小板反应蛋白基因。全长斑节对虾血小板反应蛋白(PmTSP)cDNA包含9个碱基对的5个非翻译区(UTR),2778个碱基对的开放阅读框(ORF)编码分子量为100.57 kDa的925个氨基酸的多肽和3UTR 99bp。 ScanProsite分析表明PmTSP包含四个几丁质结合II型结构域,一个EGF样结构域,八个血小板反应蛋白III型重复序列和一个血小板反应蛋白C末端结构域。通过MatGAT软件对推定的PmTSP氨基酸序列与其他已知的TSP序列进行同源性分析,结果表明PmTSP与Fennerpenaeus chinensis的序列显示出非常高的同源性(89.9%的相似性,83.8%的同一性)。对PmTSP基因的组织表达模式的分析表明,PmTSP mRNA在所有测试组织中都有表达,包括肝胰脏,卵巢,肌肉,肠,眼球中的神经分泌器官,脑,胃和心脏中的神经分泌器官,其水平最高。卵巢。此外,发现在卵巢的六个发育阶段中PmTSP表达高水平。结果表明PmTSP可能在卵巢发育中起重要作用。

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