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果蝇DnaJ同源肿瘤抑制因子Tid50的线粒体定位及时间表达

Mitochondrial localization and temporal expression of the Drosophila melanogaster DnaJ homologous tumor suppressor Tid50.

作者信息

Kurzik-Dumke U, Debes A, Kaymer M, Dienes P

机构信息

Institut für Genetik, Johannes Gutenberg-Universität, Mainz, Germany.

出版信息

Cell Stress Chaperones. 1998 Mar;3(1):12-27. doi: 10.1379/1466-1268(1998)003<0012:mlateo>2.3.co;2.

Abstract

The Drosophila melanogaster tumor suppressor gene lethal(2)tumorous imaginal discs (tid) was identified as a homolog of all dnaJ-like genes known to date which have been well preserved in evolution. Homozygous D. melanogaster l(2)tid mutants l(2)tid1, l(2)tid2 and l(2)tid3 are characterized by neoplastic transformation of the adult integumental primordia, the imaginal discs, and the death at the time of puparium formation. The first part of this study is concerned with the identification and subcellular localization of the l(2)tid-encoded protein, Tid50. The second part examines its tissue specific expression during wild-type development and in tumorous imaginal discs. To specify the function(s) of the Tid50 protein polyclonal rabbit antibodies directed against various domains of it were generated and used for staining of Western blots and whole-mounts and paraffin sections of various tissues isolated from wild-type and mutant tumor-developing animals. To identify the mutational events leading in homozygous l(2)tid mutants to abnormal expression level of l(2)tid-encoded RNA and protein, the mutant gene was isolated from homozygous l(2)tid1 and l(2)tid2 animals and sequenced.

摘要

果蝇黑腹果蝇肿瘤抑制基因致死(2)肿瘤性成虫盘(tid)被鉴定为迄今为止已知的所有dnaJ样基因的同源物,这些基因在进化过程中得到了很好的保存。纯合的黑腹果蝇l(2)tid突变体l(2)tid1、l(2)tid2和l(2)tid3的特征是成虫体壁原基、成虫盘发生肿瘤转化,并在蛹形成时死亡。本研究的第一部分涉及l(2)tid编码蛋白Tid50的鉴定和亚细胞定位。第二部分研究了其在野生型发育过程中和肿瘤性成虫盘中的组织特异性表达。为了确定Tid50蛋白的功能,制备了针对其不同结构域的多克隆兔抗体,并用于对从野生型和突变肿瘤发育动物分离的各种组织的蛋白质免疫印迹、整体标本和石蜡切片进行染色。为了鉴定导致纯合l(2)tid突变体中l(2)tid编码的RNA和蛋白质表达水平异常的突变事件,从纯合l(2)tid1和l(2)tid2动物中分离出突变基因并进行测序。

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