|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|
|WOS标题词||Science & Technology|
|类目[WOS]||Gastroenterology & Hepatology|
|研究领域[WOS]||Gastroenterology & Hepatology|
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.
|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
|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.|