Telerman A, Dodemont H, Degraef C, Galand P, Bauwens S, Van Oostveldt P, Amson R B
Institute of Interdisciplinary Research, School of Medicine, Free University of Brussels (ULB), Belgium.
Oncogene. 1992 Dec;7(12):2545-8.
A putative tumor-suppressor gene (wt1) located at chromosome 11p13 and involved in Wilms' tumor development has recently been identified as a zinc finger polypeptide-encoding gene. The purpose of this study was to characterize the protein encoded by the human wt1 gene. The region spanning the entire zinc finger domain was amplified by polymerase chain reaction (PCR) and subcloned in the pATH 3 expression vector. Polyclonal antibodies against the fused TrpE-WT protein were raised. These antibodies immunoprecipitated a 49- to 51-kDa protein from hematopoietic tumor cells labeled in vivo with [35S]methionine. Subcellular fractionation and immunohistochemistry followed by confocal microscopy indicated that the Wilms' tumor gene product (WT1) is mainly localized within the nucleus.
一个位于11号染色体p13区、与肾母细胞瘤发生相关的假定肿瘤抑制基因(wt1),最近被鉴定为一个编码锌指多肽的基因。本研究的目的是对人wt1基因编码的蛋白质进行特性分析。通过聚合酶链反应(PCR)扩增跨越整个锌指结构域的区域,并亚克隆到pATH 3表达载体中。制备了针对融合的TrpE-WT蛋白的多克隆抗体。这些抗体从体内用[35S]甲硫氨酸标记的造血肿瘤细胞中免疫沉淀出一种49至51 kDa的蛋白质。亚细胞分级分离和免疫组织化学,随后进行共聚焦显微镜检查表明,肾母细胞瘤基因产物(WT1)主要定位于细胞核内。