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Recovery of recombinant zebrafish p53 protein from inclusion bodies and its binding activity to p53 mRNA in vitro
Zhao, Xiangzhong1,2; Liu, Ming1; Wu, Ning1; Ding, Lili1; Liu, Haizhou1; Lin, Xiukun1
2010-08-01
Source PublicationPROTEIN EXPRESSION AND PURIFICATION
Volume72Issue:2Pages:262-266
SubtypeArticle
Abstractp53 protein is an important regulation factor that can bind to p53 mRNA to regulate its translation in human and murine. To determine if a similar interaction exists in zebrafish and if the interaction affects zebrafish development, we cloned and expressed p53 protein from zebrafish in Escherichia coli. Soluble p53 protein with high purity was successfully obtained using the optimized renaturation approach. Results of a UV-crosslinking experiment and immunoprecipitation:RT-PCR analysis confirmed that the purified p53 protein could bind specifically to its cognate mRNA. Our results suggest that selecting a suitable buffer is important for renaturing p53 protein from inclusion bodies. We also demonstrated a specific interaction between p53 and it own RNA in zebrafish. Measurement of the binding activity may be a useful approach for identifying the activity of recombinant p53 protein in vitro. (C) 2010 Elsevier Inc. All rights reserved.
KeywordP53 Zebrafish Rna Binding Protein Protein Expression Inclusion Body
DOI10.1016/j.pep.2010.03.028
Indexed BySCI
Language英语
WOS IDWOS:000278193200017
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Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://ir.qdio.ac.cn/handle/337002/32772
Collection实验海洋生物学重点实验室
Affiliation1.Chinese Acad Sci, Inst Oceanol, Qingdao 266071, Peoples R China
2.Chinese Acad Sci, Grad Sch, Beijing 100039, Peoples R China
First Author AffilicationInstitute of Oceanology, Chinese Academy of Sciences
Recommended Citation
GB/T 7714
Zhao, Xiangzhong,Liu, Ming,Wu, Ning,et al. Recovery of recombinant zebrafish p53 protein from inclusion bodies and its binding activity to p53 mRNA in vitro[J]. PROTEIN EXPRESSION AND PURIFICATION,2010,72(2):262-266.
APA Zhao, Xiangzhong,Liu, Ming,Wu, Ning,Ding, Lili,Liu, Haizhou,&Lin, Xiukun.(2010).Recovery of recombinant zebrafish p53 protein from inclusion bodies and its binding activity to p53 mRNA in vitro.PROTEIN EXPRESSION AND PURIFICATION,72(2),262-266.
MLA Zhao, Xiangzhong,et al."Recovery of recombinant zebrafish p53 protein from inclusion bodies and its binding activity to p53 mRNA in vitro".PROTEIN EXPRESSION AND PURIFICATION 72.2(2010):262-266.
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