Trypanosoma brucei: molecular cloning of homologues of small GTP-binding proteins involved in vesicle trafficking.

Trypanosoma brucei: molecular cloning of homologues of small GTP-binding proteins involved in vesicle trafficking.
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布氏锥虫:参与囊泡运输的小 GTP 结合蛋白同源物的分子克隆。

DOI:
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发表时间:
1995
影响因子:
2.1
通讯作者:
J. Boothroyd
J. Boothroyd
中科院分区:
医学4区
文献类型:
--
作者:
Mark C. Field;J. Boothroyd

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GTPases rab 亚家族的成员被认为是真核生物囊泡运输的重要组成部分。单个 rab 蛋白具有显着程度的特异性亚细胞定位。作为研究原生动物寄生虫布氏锥虫中这些蛋白质的第一个方法,我们使用基于简并聚合酶链反应的策略对 rab 亚科进行了调查,该策略利用了微型外显子和高度保守的 WDTAGQE 盒(包含部分 GTP 结合位点)。从前环阶段 cDNA 中获得了许多布氏锥虫的部分 cDNA,其中 7 个克隆(命名为 rtb 1、3-7、9)显然是 rab 家族的成员。 Poly(A) 选择的 RNA 的 Northern 分析表明,rtb1、3、4 和 7 在 T. brucei 的两个生命阶段均以低水平组成型表达。通过对锥虫基因组 DNA 进行 Southern 分析以及对分离的基因组克隆进行特异性 PCR,我们发现其中两个基因 RTB1 和 RTB7 在 T. brucei 基因组中相邻或至少紧密相连,而其他五个基因则分散。这些数据为剖析该生物体中不寻常的分泌途径提供了重要的分子试剂。
Members of the rab subfamily of GTPases have been implicated as important components in vesicle trafficking in the eukaryotes. Individual rab proteins have a remarkable degree of specific subcellular localization. As a first approach to the study of these proteins in the protozoan parasite Trypanosoma brucei we have undertaken a survey of the rab subfamily using a strategy based on degenerate polymerase chain reaction utilizing the miniexon and the highly conserved WDTAGQE box which comprises part of the GTP-binding site. A number of T. brucei partial cDNAs were obtained from procyclic stage cDNA, and seven of these clones (designated rtb 1, 3-7, 9) were clearly members of the rab family. Northern analysis of poly(A)-selected RNA indicates that rtb1, 3, 4, and 7 are constitutively expressed at low levels in both life-stages of T. brucei. By Southern analysis of trypanosome genomic DNA and specific PCR from an isolated genomic clone, we show that two of the genes, RTB1 and RTB7, are adjacent or at least closely linked in the T. brucei genome, while the other five are dispersed. These data provide important molecular reagents for dissecting the unusual secretory pathway in this organism.
DOI: 10.1242/jcs.105.4.1101
发表时间: 1993
影响因子: 4
作者:
Bangs,JD;Uyetake,L;Brickman,MJ;Balber,AE;Boothroyd,JC
通讯作者: Boothroyd,JC
锥虫变异表面糖蛋白基因家族的基因组组织和背景。
DOI: 10.1016/0022-2836(92)90097-4
发表时间: 1992
影响因子: 5.6
作者:
Beals,TP;Boothroyd,JC
通讯作者: Boothroyd,JC
多基因转录物是锥虫中钙调蛋白 mRNA 的前体。
DOI: 10.1002/j.1460-2075.1988.tb02833.x
发表时间: 1988
期刊: The EMBO journal
影响因子: --
作者:
Tschudi,C;Ullu,E
通讯作者: Ullu,E