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中国生物工程杂志

China Biotechnology
China Biotechnology  2009, Vol. 29 Issue (03): 36-40    DOI:
    
Site-directed Mutagenesis, Expression And Purification Of PZR
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Abstract  

Objective In order to express and purify the mono-phosphorylated PZR proteins. Methods The bi-- phosphorylated His-PZR recombined prokaryotic expressions plasmids was used as the template, then site-directed mutagenesis was used to amplified the mono- phosphorylated PZR a Tyr263(TAT) Phe263(TTT)and PZR b Tyr241(TAT) Phe241(TTT) mutants. The mutants’ and C-Src recombined prokaryotic expressions plasmids were co-transformed to the E.Coli. BL21. Through IPTG induction and Ni-NTA purification, we want to get the mono-phosphorylated PZR a and PZR b proteins. Results The mono- phosphorylated PZR a and PZR b recombined prokaryotic expressions plasmids have successfully been constructed; The mutants have successfully been induced to express by IPTG. They represent 8.5% of the total bacterial protein in E.Coli. BL21, and theirs relative molecule mass is 10 000. After Ni-NTA purification, the mono-phosphorylated PZR a and PZR b proteins have been obtained. Conclusion These results laid a foundation to future study the bio-function mechanism between different ITIM of PZR with protein tyrosine phosphatase in the signal transduction.



Key wordsPZR;ITIM;Site-directed mutation;Expression;Purification     
Received: 13 October 2008      Published: 31 March 2009
ZTFLH:  R392.11  
Cite this article:

MO Yi- Wang-Wei- Liang-Fang-Fang- Yang-Xu-Fen- Song-Qian- Jiang-He-Sheng- Wayne Zhou. Site-directed Mutagenesis, Expression And Purification Of PZR. China Biotechnology, 2009, 29(03): 36-40.

URL:

https://manu60.magtech.com.cn/biotech/     OR     https://manu60.magtech.com.cn/biotech/Y2009/V29/I03/36

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