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旋毛虫一些热诱导基因的鉴定。

Identification of some heat-induced genes of Trichinella spiralis.

作者信息

Mak C H, Su K W, Ko R C

机构信息

Department of Zoology, The University of Hong Kong, Pokfulam, China.

出版信息

Parasitology. 2001 Sep;123(Pt 3):293-300. doi: 10.1017/s0031182001008320.

Abstract

Three heat-induced genes of the infective-stage larvae of Trichinella spiralis were successfully identified by the suppression subtractive hybridization (SSH) technique. As indicated by reverse Northern blotting, 19 of 25 clones were scored as differentially transcribed in the heat-shocked infective-stage larvae. The sequencing data showed the presence of 12 different genes. Three were homologous to histone H3, histone H2B and translationally controlled tumour protein (TCTP). A 0.6 kb cDNA of histone H3 was generated by the RACE method and sequenced. It contained an open reading frame of 136 amino acids that demonstrated 94% identity with genes from Drosophila hydei. Semi-quantitative RT-PCR indicated that after heat-shock treatment, the expression levels of histone H3, histone H2B and TCTP increased 4.8, 27 and 5.7-fold, respectively. Northern analysis confirmed the upregulation of histone H3, histone H2B and TCTP transcripts. The upregulation of these genes during stress conditions has not been reported in parasitic organisms. The stress proteins may play an active role to sustain the parasite after exposure to hostile host factors.

摘要

利用抑制性消减杂交(SSH)技术成功鉴定了旋毛虫感染期幼虫的三个热诱导基因。反向Northern印迹分析表明,25个克隆中有19个在热激感染期幼虫中被判定为差异转录。测序数据显示存在12个不同的基因。其中三个与组蛋白H3、组蛋白H2B和翻译控制肿瘤蛋白(TCTP)同源。通过RACE方法获得了组蛋白H3的0.6 kb cDNA并进行了测序。它包含一个136个氨基酸的开放阅读框,与海德氏果蝇的基因具有94%的同一性。半定量RT-PCR表明,热激处理后,组蛋白H3、组蛋白H2B和TCTP的表达水平分别增加了4.8倍、27倍和5.7倍。Northern分析证实了组蛋白H3、组蛋白H2B和TCTP转录本的上调。在寄生虫中尚未报道这些基因在应激条件下的上调情况。应激蛋白可能在寄生虫暴露于宿主有害因素后发挥积极作用以维持其生存。

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