日本沼虾mu型可溶性谷胱甘肽S-转移酶的克隆与表达
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国家科技部“863”项目(2007AA09Z437)和国家自然科学基金项目(30670367)


Molecular cloning and in vivo expression analysis of mu-class soluble glutathione S-transferase in shrimp 
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    摘要:

    利用简并引物从日本沼虾肝脏克隆mu型sGST基因cDNA核心片段获得其sGST氨基酸序列。序列分析表明日本沼虾肝脏mu型sGST基因cDNA核心序列长299 bp,编码99个氨基酸。日本沼虾sGST与南美白对虾(Litopenaeus vannamei)、小鼠(Mus musculus)、大鼠(Rattus norvegicus)、肩突硬蜱(Ixodes scapularis)、扇头蜱(Rhipicephalus appendiculatus)、热带爪蟾(Xenopus tropicalis)、微小牛蜱(Rhipicephalus microplus)和太平洋牡蛎(Crassostrea gigas)的mu型sGST基因核苷酸同源性为60%左右,表明所克隆的日本沼虾sGST亦属于mu型sGST。通过对日本沼虾活体浸泡MC-LR,发现微囊藻毒素对日本沼虾肝脏mu型sGST基因表达没有显著的诱导作用。

    Abstract:

    Soluble glutathione S-transferase (sGST) is extremely important for metabolic detoxication by catalyzing the conjugation of GSH (glutathione) with microcystins.In this study,the amino acid sequence of a mu-class sGST cDNA fragment from the liver of Macrobrachium nipponensis were firstly obtained by RT-PCR using degenerated primers.Sequence analysis revealed that the cloned mu-class sGST cDNA fragment was 299 bp in length,encoding 99 amino acids.Homologous analysis showed that the nucleotide sequence of the glutathione S-transferase gene in M.nipponensis was 60% homologus to the glutathione S-transferase gene of Litopenaeus vannamei,Rattus norvegicus,Mus musculus,Ixodes scapularis,Rhipicephalus appendiculatus,Xenopus tropicalis,Rhipicephalus microplus and Crassostrea gigas.This suggested that the sGST cloned in this study belonged to mu-class GST.The result of further experiment showed that mu-class sGST level did not change significantly in M.nipponensis exposed in MC-LR.

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瞿春梅,梁旭方,张进,何珊,沈丹.日本沼虾mu型可溶性谷胱甘肽S-转移酶的克隆与表达[J].华中农业大学学报,2013,32(2):103-108

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