Transcriptional analyses of Trypanosoma brucei gambiense from infected mice and in vitro culture

Noritaka Kuboki, Michael K. Kibe, O. Thekisoe, C. Sugimoto, N. Inoue
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Abstract

ABSTRACT With the hypothesis that African trypanosomes could have in vivo specific genes for adaptation to host’s environment, the present study was conducted by using suppressive subtractive hybridization (SSH) technique to seek the highly expressed genes especially in host. A total of 328 clones from the in vivo SSH library and that of 160 clones from the in vitro SSH library were analyzed in order to determine their expression levels, but none of the above-mentioned genes showed differential expression. This indicates that no trypanosome genes could be differentially expressed either the in vivo or in vitro propagated trypanosomes. Alternatively, there might be limitation for detecting specifically expressed genes in African trypanosomes using this method, because of their polycistronic gene expression. Key Words : differential expression; infection; Trypanosoma brucei The development of axenic culture methods for both bloodstream form (BSF) and procyclic form (PCF) of African trypanosomes (Hirumi and Hirumi 1989; Hirumi and Hirumi 1991) has contributed to further investigation of African trypanosomes. However, this invention has given rise to a question of whether biological characteristics of
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感染小鼠及体外培养的布氏冈比亚锥虫转录分析
摘要本研究假设非洲锥虫体内可能存在适应宿主环境的特异性基因,采用抑制减法杂交(suppression subtractive hybridization, SSH)技术寻找在宿主体内高表达的基因。我们对体内SSH文库中的328个克隆和体外SSH文库中的160个克隆进行了分析,以确定它们的表达水平,但上述基因均未出现差异表达。这表明在体内和体外繁殖的锥虫均不存在基因差异表达。另外,由于非洲锥虫的多顺反子基因表达,使用这种方法检测特异性表达基因可能存在局限性。关键词:微分表达式;感染;非洲锥虫血流型(BSF)和顺环型(PCF)的无菌培养方法的发展(Hirumi and Hirumi 1989;Hirumi和Hirumi(1991)对非洲锥虫的进一步研究做出了贡献。然而,本发明引起了是否生物特性的问题
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