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首页> 外文期刊>Molecular and Cellular Biology >Transient activity assays of the Trypanosoma brucei variant surface glycoprotein gene promoter: control of gene expression at the posttranscriptional level.
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Transient activity assays of the Trypanosoma brucei variant surface glycoprotein gene promoter: control of gene expression at the posttranscriptional level.

机译：布鲁氏锥虫变体表面糖蛋白基因启动子的瞬时活性测定：转录后水平的基因表达控制。

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The putative promoter of the variant surface glycoprotein (VSG) gene of Trypanosoma brucei was cloned into a plasmid containing the chloramphenicol acetyltransferase (CAT) gene. After electroporation into trypanosomes, this construct directed the expression of the CAT reporter gene. The essential region for promoter activity was found to reside within 88 bp upstream of the putative transcription start site. Transcription of the CAT construct occurred at approximately the same level in both bloodstream and procyclic forms and was resistant to alpha-amanitin. However, CAT expression appeared to be modulated in the two forms of the parasite. Sequences 3' to the gene seemed to be important in this respect, as CAT activity in bloodstream forms was readily detectable only when the 3' region of a VSG cDNA was placed downstream of the CAT gene. Two separate VSG gene promoter sequences, both cloned from T. brucei AnTat 1.3A, were equally able to direct CAT expression, which suggests that there are a number of potential VSG gene promoters in the genome, although usually only one expression site is fully active at any one time.
机译：将布鲁氏锥虫变体表面糖蛋白（VSG）基因的推定启动子克隆到含有氯霉素乙酰基转移酶（CAT）基因的质粒中。电穿孔入锥虫体后，该构建体指导CAT报告基因的表达。发现启动子活性的必要区域位于推定转录起始位点上游88 bp之内。 CAT构建体的转录在血流和前环形式中都以大约相同的水平发生，并且对α-amanitin具有抗性。但是，CAT表达似乎以两种形式的寄生虫被调节。在这方面，该基因的3'序列似乎很重要，因为只有将VSG cDNA的3'区放置在CAT基因的下游时，才能轻易检测到血流形式的CAT活性。分别从布氏锥虫AnTat 1.3A克隆的两个单独的VSG基因启动子序列同样能够指导CAT表达，这表明基因组中有许多潜在的VSG基因启动子，尽管通常只有一个表达位点是完全活跃的在任何时候。

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