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首页> 外文期刊>Nucleic acids research >Faithful expression of a heterologous gene carried on an artificial macronuclear chromosome in Euplotes crassus
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Faithful expression of a heterologous gene carried on an artificial macronuclear chromosome in Euplotes crassus

机译：真实地表达了人工大核染色体上携带的异源基因

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摘要

Macronuclear chromosomes of hypotrichous ciliates are gene-sized molecules carrying the coding sequence flanked by short non-translated regions and bounded by telomeres. We have constructed artificial chromosomes for investigation of transcription in the macronucleus of Euplotes crassus. The neo gene was put under the control of the 5′-non-translated region of the TBP gene of E.crassus. These molecules were introduced into the cell with the help of liposomes. The cells were transformed and survived high concentrations of geneticin. The artificial chromosomes were kept in the macro-nucleus for at least 50 days at a copy number of about 200 per macronucleus. Expression of the gene was shown by reverse transcription of the neo messenger. The transcription start was mapped and found to coincide with that found on the natural macronuclear chromosome encoding TBP in E.crassus.
机译：纤毛虫纤毛虫的大核染色体是基因大小的分子，带有编码序列，其侧翼是短的非翻译区，并由端粒限制。我们已经构建了人工染色体，用于研究真蟹真核大核中的转录。 neo基因被置于E.crassus的TBP基因的5'-非翻译区的控制之下。这些分子借助于脂质体被引入细胞。细胞被转化并在高浓度的遗传霉素中存活。人工染色体在大核中保持至少50天，每个大核的拷贝数约为200。该基因的表达通过新信使的逆转录显示。对转录起点进行定位，发现其与在crassus中编码TBP的天然大核染色体上发现的一致。

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