Characterization and disruption of a new Trypanosoma brucei repetitive flagellum protein, using double-stranded RNA inhibition
Characterization and disruption of a new Trypanosoma brucei repetitive flagellum protein, using double-stranded RNA inhibition
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DOI：
10.1016/s0166-6851(00)00319-4
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发表时间：
2000-12-01
影响因子：
1.5
通讯作者：
Baltz, T
中科院分区：
文献类型：
--
作者：
Bringaud, F;Robinson, DR;Baltz, T
在布鲁氏锥虫中，我们克隆了一个类似于葡萄糖转运基因簇下游5 kb的基因，包含可变数量的102 bp重复序列。该基因编码的蛋白质在数据库中没有同源物。针对34个氨基酸重复基序识别蛋白的抗体，范围从145到270 kDa不等，取决于菌株，在血流和顺环形式的布鲁氏体中。检测到的蛋白质的表观分子质量与34个氨基酸重复数（3 ~ 40）之间存在相关性。我们称这种蛋白为鞭毛过渡区成分（FTZC），因为它位于过渡区内轴突的近端区域。FTZC是唯一报道的在过渡区存在锥虫体蛋白的例子。为了确定FTZC的作用，我们开发了一种基于双链RNA条件表达的基因失活新策略。在四环素存在的情况下，双链RNA的表达，我们观察到FTZC在etro1125和etro427菌株中完全消失。在使用的生长条件下，FTZC的分子消融不会产生任何明显的表型，如致命性、生长速度或细胞形状的改变。(C) 2000 Elsevier Science B.V.版权所有
In Trypanosoma brucei, we have cloned a gene similar to 5 kb downstream of the glucose transporter gene cluster, containing a variable number of 102 bp repeats. This gene encodes a protein with no homologues in the data bases. Antibodies raised against the 34 amino acids repeated motif recognized proteins ranging from 145 to 270 kDa, depending on strains, in both bloodstream and procyclic forms of T. brucei. A correlation was established between the apparent molecular mass of the detected proteins and the number of 34 amino acid repeats which varies from 3 to 40. We have called this protein the flagellum transition zone component (FTZC) due to its localization to the proximal region of the axoneme, within the transition zone. FTZC is the only reported example of a trypanosomal protein present in the transition zone. To determine the role of FTZC we developed a new strategy of gene inactivation based on conditional expression of double-stranded RNA. In the presence of tetracycline, expression of the double-stranded RNA, we observed a complete disappearance of FTZC in the EATRO 1125 and EATRO 427 strains of T. brucei. Molecular ablation of FTZC does not generate any obvious phenotype such as, lethality, modification of growth rate or cellular shape, in the growth conditions used. (C) 2000 Elsevier Science B.V. All rights reserved.