IR@PKUHSC  > 北京大学临床肿瘤学院
学科主题临床医学
Screening and identification of proteins interacting with nucleostemin
Yang, Hai-Xia1,2; Jin, Geng-Lin1,2; Meng, Ling1,2; Zhang, Jian-Zhi1,2; Liu, Wen-Bin1,2; Shou, Cheng-Chao1,2
关键词Nucleostemin Yeast Two-hybrid Co-ip
刊名WORLD JOURNAL OF GASTROENTEROLOGY
2005-08-21
11期:31页:4812-4814
收录类别SCI
文章类型Article
WOS标题词Science & Technology
类目[WOS]Gastroenterology & Hepatology
研究领域[WOS]Gastroenterology & Hepatology
英文摘要

AIM: To identify the proteins interacting with nucleostemin (NS), thereby gaining an insight into the function of NS.

METHODS: Yeast two-hybrid assay was performed to screen a human placenta cDNA library with the full length of NS as a bait. X-Gal assay and beta-galactosidase filter assay were subsequently conducted to check the positive clones and the gene was identified by DNA sequencing. To further confirm the interaction of two proteins, the DNA fragment coding NS and the DNA fragment isolated from the positive clone were inserted into the mammalian expression vector pcDNA3 and pcDNA3-myc, respectively. Then, two plasmids were cotransfected into the COS-7 cells by DEAE-dextron. The total protein from the cotransfected cells was extracted and coimmunoprecipitation and Western blot were performed with suitable antibodies sequentially.

RESULTS: Two positive clones that interacted with NS were obtained from human placenta cDNA library. One was an alpha isoform of human protein phosphatase 2 regulatory subunit B (B56) (PPP2R5A) and the other was a novel gene being highly homologous to the gene associated with spondylo paralysis. The co-immunoprecipitation also showed that NS specifically interacted with PPP2R5A.

CONCLUSION: NS and PPP2R5A interact in yeast and mammalian cells, respectively, which is helpful for addressing the function of NS in cancer development and progression. (c) 2005 The WJG Press and Elsevier Inc. All rights reserved.

语种英语
WOS记录号WOS:000208317800007
项目编号200BA711A11A06 ; H020220020310
资助机构National High Technology Research and Development Program of China ; Beijing Science and Technology Project
引用统计
被引频次:11[WOS]   [WOS记录]     [WOS相关记录]
文献类型期刊论文
条目标识符http://ir.bjmu.edu.cn/handle/400002259/54868
专题北京大学临床肿瘤学院
北京大学临床肿瘤学院_胸部肿瘤外二科
北京大学临床肿瘤学院_生化与分子生物学研究室
作者单位1.Beijing Inst Canc Res, Beijing 100034, Peoples R China
2.Peking Univ, Sch Oncol, Dept Biochem & Mol Biol, Beijing 100034, Peoples R China
推荐引用方式
GB/T 7714
Yang, Hai-Xia,Jin, Geng-Lin,Meng, Ling,et al. Screening and identification of proteins interacting with nucleostemin[J]. WORLD JOURNAL OF GASTROENTEROLOGY,2005,11(31):4812-4814.
APA Yang, Hai-Xia,Jin, Geng-Lin,Meng, Ling,Zhang, Jian-Zhi,Liu, Wen-Bin,&Shou, Cheng-Chao.(2005).Screening and identification of proteins interacting with nucleostemin.WORLD JOURNAL OF GASTROENTEROLOGY,11(31),4812-4814.
MLA Yang, Hai-Xia,et al."Screening and identification of proteins interacting with nucleostemin".WORLD JOURNAL OF GASTROENTEROLOGY 11.31(2005):4812-4814.
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