Heterologous expression of a plant arginine decarboxylase gene in Trypanosoma cruzi.
Heterologous expression of a plant arginine decarboxylase gene in Trypanosoma cruzi.
复制标题
植物精氨酸脱羧酶基因在克氏锥虫中的异源表达。
DOI：
10.1016/j.bbagen.2004.06.017
复制
发表时间：
2004
期刊：
影响因子：
--
通讯作者：
Algranati,IsraelD
中科院分区：
文献类型：
--
作者：
Carrillo,Carolina;Serra,MaríaP;Pereira,ClaudioA;Huber,Alejandra;González,NélidaS;Algranati,IsraelD
野生型克氏锥虫表皮马鞭毛虫缺乏精氨酸脱羧酶（ADC）酶活性。然而，用含有燕麦ADC cDNA编码区的重组质粒转化这些寄生虫会导致该酶的短暂异源表达，这表明缺乏内源性机制来抑制假设的自身ADC基因的表达或用于测量其酶活性的检测。通过产物鉴定、催化反应的化学计量学、α-二氟甲基精氨酸（DFMA）的特异性抑制及其代谢转换研究，对外源ADC酶在转基因克氏体中的表达进行了表征。异源ADC在克氏锥虫体内的半衰期约为150 min。生物信息学研究和聚合酶链反应（PCR）分析似乎表明野生型克氏T.基因组中缺乏adc样DNA序列。
Wild-type Trypanosoma cruzi epimastigotes lack arginine decarboxylase (ADC) enzymatic activity. However, the transformation of these parasites with a recombinant plasmid containing the oat ADC cDNA coding region gave rise to the transient heterologous expression of the enzyme, suggesting the absence of endogenous mechanisms that could inhibit the expression of a hypothetical own ADC gene or the assay used to measure its enzymatic activity. The foreign ADC enzyme expressed in the transgenic T. cruzi was characterized by identification of the products, the stoichiometry of the catalysed reaction, the specific inhibition by α-difluoromethylarginine (DFMA) and the study of its metabolic turnover. The half-life of the heterologous ADC activity in T. cruzi was about 150 min. Bioinformatics studies and polymerase chain reaction (PCR) analyses seem to indicate the absence of ADC-like DNA sequences in the wild-type T. cruzi genome.