Expression of recombinant 32 000 doliton polypeptide of mouse enamelin in Escherichia coli and its purification
10.3760/cma.j.issn.1002-0098.2011.s1.003
- VernacularTitle:小鼠重组釉蛋白32000多肽的原核表达和纯化
- Author:
Ping L(U)
1
;
Hua TIAN
;
Xue-Jun GAO
Author Information
1. 北京大学口腔医学院(口腔医院)
- Keywords:
Gene expression;
Enamelin;
Purification
- From:
Chinese Journal of Stomatology
2011;46(z1):11-14
- CountryChina
- Language:Chinese
-
Abstract:
Objective To examine the expression of the recombinant mouse 32 000 enamelin in Escherichia coli(Ec).Methods The desired 32 000 enamelin cDNA fragment was obtained by polymerase chain reaction (PCR).The segment was inserted into pGEM-T Easy vector and transformened into Ec JM109.The positive clone was analyzed by restriction endonuclease mapping and DNA sequencing.The 312 bp aimed fragment was inserted into pGEX-4T1 vector and the recombinant protein was induced by isopropyl thiogalactoside in Ec BL-21.Results The recombinant expressive plasmid pGEX-4T1-EN312 was successfully constructed.After induced with isopropythio-galactoside,a new fusion protein band near Mr 47 ×103 appeared on SDS polyacrylamide gel electrophoresis.Conclusions pGEX-4T1-EN312-BL21 system is used successfully to express and purify recombinant mouse 32 000 enamelin polypeptide.
