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MnHSP90 cDNA 基因的特征及其在日本沼虾(Macrobrachium nipponense)卵巢发育过程中的表达。

MnHSP90 cDNA characterization and its expression during the ovary development in oriental river prawn, Macrobrachium nipponense.

机构信息

Department of Biology, East China Normal University, 200062, Shanghai, People's Republic of China.

出版信息

Mol Biol Rep. 2011 Feb;38(2):1399-406. doi: 10.1007/s11033-010-0243-7. Epub 2010 Jul 31.

Abstract

Heat shock protein 90 (HSP90) is not only involved in environmental stress but also plays roles in the ovary development in some vertebrates. To understand its role in crustacean, we examined the HSP90 cDNA for the first time in the ovary and hepatopancreas of the oriental river prawn, Macrobrachium nipponense and designated this protein as MnHSP90 in this study. The MnHSP90 was cloned by the methods of degenerated oligonucleotide primers and rapid amplification of the cDNA ends (RACE). Bioinformatics analysis showed that the MnHSP90 cDNA was 2,684 bp in length, containing a 126 bp 5' untranslated region (UTR), a 359 bp 3' UTR, and an open reading frame (ORF) of 2,199 bp encoding a 732-amino acid polypeptide with predicted molecular mass of 84.3 KDa. Sequence alignment showed that the MnHSP90 shared 72-79% identity with other animals. Real-time quantitative PCR (qPCR) analysis demonstrated that the MnHSP90 mRNA was ubiquitously detected in all tested tissues, with the highest expression in the thoracic ganglia, the mediate in heart, muscle and intestine, and the lowest in haemocytes and gills. The MnHSP90 mRNA levels in the hepatopancreas and ovary of M. nipponense reached a maximum at the stage III (early vitellogenic stage) and stage IV (later vitellogenic stage) ovaries, respectively, and then decreased significantly in both tissues as the ovarian development proceeded. The level of MnHSP90 expression in the hepatopancreas was higher than that in the ovary when compared with in the same ovarian developmental stage. Our results indicate that MnHSP90 is involved in ovarian development in oriental river prawn and may play a regulatory role in ovary maturation.

摘要

热休克蛋白 90(HSP90)不仅参与环境应激反应,还在一些脊椎动物的卵巢发育中发挥作用。为了了解其在甲壳动物中的作用,我们首次在日本沼虾(Macrobrachium nipponense)的卵巢和肝胰腺中检测到 HSP90 cDNA,并在本研究中将该蛋白命名为 MnHSP90。通过简并寡核苷酸引物和快速扩增 cDNA 末端(RACE)方法克隆 MnHSP90。生物信息学分析表明,MnHSP90 cDNA 长 2684 bp,包含 126 bp 的 5'非翻译区(UTR)、359 bp 的 3'UTR 和一个 2199 bp 的开放阅读框(ORF),编码一个 732 个氨基酸的多肽,预测分子量为 84.3 kDa。序列比对表明,MnHSP90与其他动物的同源性为 72-79%。实时定量 PCR(qPCR)分析表明,MnHSP90 mRNA 在所有检测组织中均有表达,在胸神经节中表达最高,在心、肌肉和肠中中等表达,在血细胞和鳃中表达最低。MnHSP90 mRNA 在日本沼虾肝胰腺和卵巢中的表达水平在卵巢 III 期(早期卵黄发生期)和 IV 期(晚期卵黄发生期)达到最高,然后在两个组织中均显著下降随着卵巢发育的进行。与同一卵巢发育阶段相比,MnHSP90 在肝胰腺中的表达水平高于卵巢。我们的结果表明,MnHSP90 参与了日本沼虾的卵巢发育,可能在卵巢成熟过程中发挥调节作用。

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