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抗果蝇paf1复合体抗体的制备及其特性

魏文祥, 杨吉成, 庄文卓, 白艳艳, 盛伟华, 缪竞诚

苏州大学医学院细胞与分子生物学教研室.江苏,苏州 215123

摘要

Paf1复合体己在酵母菌被发现，并阐明了其在转录及RNA加工过程中发挥功能。作者在果蝇基因组数据库中鉴定出paf1的三个同源成分：paf1，CDC73和RTF1。应用纯化的重组蛋白，作者制备了抗果蝇paf1，CDC73和RTF1的抗体。这些抗体不但能够检测出果蝇内源性paf1成分，而且能检测出人类的paf1复合体的亚基。在多线染色体上，这些抗体所检测到的paf1复合体均位于转录的活性位点，并且与磷酸化的RNA聚合酶重合，表明果蝇paf1复合体参与了转录及其相关的分子活动。

关键词： paf1复合体; paf1; CDC73; RTF1; 转录

Generation and characterization of antibody against paf1 complex in {sl Drosophila melanogaster}

Wei Wenxiang, Yang Jicheng, Zhuang Wenzhuo, Bai Yanyan, Sheng Weihua, Miao Jingcheng

Department of Cellular and Molecular Biology, School of Medicine, Soochow University.Suzhou 215123,Jiangsu

Abstract

Paf1 complex was identified in yeast and characterized to function in transcription and its related events. We identified the Drosophila homological components of pafl, CDC73 and RTF1 of paf1 complex. The genes encoding Drosophila pafl, CDC73 and RTF1 were cloned and expressed. With the purified recombinant proteins of truncated components of paf1 complex, antibodies against the Drosophila pafl, CDC73 and RTF1 were generated. These antibodies have been shown to be able to detect the endogenous pafl subunits as well as their human counterparts in the HeLa extract. On Drosophila polytene chromosomes, these antibodies have been demonstrated to locate the paf1 complex at actively transcribing sites, which co-localized with phosphorylated RNA polymerase Ⅱ, indicating that paf1 complex in Drosophila is involved in transcription or the events coupling with transcription.

Key words: pafl complex;paf1;CDC73;RTF1;transcription

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引用本文：

魏文祥, 杨吉成, 庄文卓, 白艳艳, 盛伟华, 缪竞诚. 抗果蝇paf1复合体抗体的制备及其特性[J]. 生理学报 2006; 58 (6): .

Wei Wenxiang, Yang Jicheng, Zhuang Wenzhuo, Bai Yanyan, Sheng Weihua, Miao Jingcheng. Generation and characterization of antibody against paf1 complex in {sl Drosophila melanogaster}. Acta Physiol Sin 2006; 58 (6): (in Chinese with English abstract).