This work was partly supported by National Natural Science Foundation of Jiangsu Province (No. BK2005028) and National Natural Science Foundation of China (No. 30140007).

 

Generation and characterization of antibody against paf1 complex of Drosophila melanogaster

 

WEI Wen-Xiang1,2, YANG Ji-Cheng1, ZHUANG Wen-Zhuo1, BAI Yan-Yan Bai1, SHENG Wei-Hua1, MIAO Jing-Cheng1,*

1Department of Cellular and Molecular Biology School of Medicine, Suzhou University, Suzhou 215123, China; 2Howard Hughes Medical Institute, Division of Nucleic Acids Enzymology, Department of Biochemistry,University of Medicine and Dentistry of New Jersey, Robert Wood Johnson Medical School, Piscataway, New Jersey 08854, USA

 

Abstract: Paf1 complex was identified in yeast and characterized to function in transcription and its related events. We identified the Drosophila homolog components of paf1, CDC73 and RTF1 of paf1 complex. The genes encoding Drosophila paf1, CDC73 and RTF1 were cloned and expressed. With the purified recombinant proteins of truncated components of paf1 complex, antibodies against the Drosophila paf1, CDC73 and RTF1 were generated. These antibodies have been shown to be able to detect the endogenous paf1 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: paf1 complex; paf1; CDC73; RTF1; transcription

 

 

抗果蝇 paf1 复合体抗体的制备及其特性

 

魏文祥1,2，杨吉成1，庄文卓1，白艳艳1，盛伟华1，缪竞诚1,*

苏州大学医学院细胞与分子生物学教研室，苏州 215123

 

摘 要：Paf1复合体已在酵母菌获得发现和阐明，其在转录及RNA的处理中发挥功能。我们在果蝇中鉴定出Paf1的三个同类成分：paf1, CDC73 和 RTF1。应用纯化的重组蛋白，我们制备了抗果蝇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

中图分类号：R3