PCR-based Gene Synthesis, Molecular Cloning, High Level Expression, Purification, and Characterization of Novel Antimicrobial Peptide, Brevinin-2R, in Escherichia Coli

2007 ◽  
Vol 149 (2) ◽  
pp. 109-118 ◽  
Author(s):  
Faramarz Mehrnejad ◽  
Hossein Naderi-Manesh ◽  
Bijan Ranjbar ◽  
Bahman Maroufi ◽  
Ahmad Asoodeh ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Peptide ◽  
Molecular Cloning ◽  
Gene Synthesis ◽  
Purification And Characterization ◽  
High Level Expression ◽  
High Level

Expression, Purifi cation, and Characterization of Functional Recombinant Human Daintain/AIF-1 in Escherichia coli

2010 ◽  
Vol 65 (11-12) ◽  
pp. 726-732 ◽  
Author(s):  
Wei Wang ◽  
Bin Huang ◽  
Zhi-Guo Feng ◽  
Xiao-Ping Chen ◽  
Win-Xin Tang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Purification And Characterization ◽  
High Level Expression ◽  
Expression Plasmid ◽  
Recombinant Peptide ◽  
Increase Blood Glucose ◽  
Sds Page ◽  
High Level ◽  
Mcf 7

Daintain/AIF-1 was identified from injured rat carotid arteries and porcine intestine in the mid 1990s. It is involved in autoimmune disorders, chronic rejection of allografts, gliomas, and breast cancer. Since it is convenient and economical to obtain such a peptide biologically, in this study, we describe the expression, purification, and characterization of recombinant human daintain/AIF-1 (rhdaintain/AIF-1). The backbone of vector pET32a, a high-level expression plasmid, was used to construct the pET32a-daintain/AIF-1 plasmid for daintain/AIF-1 expression in Escherichia coli. The recombinant daintain/AIF-1 protein was solubly expressed in the BL21 (DE3) strain and was purifi ed by Ni2+ affinity chromatography. After purification, the recombinant protein showed the expected size of 18 kDa on Tricine-SDS-PAGE gels which was further confirmed by Western blotting. A total of 34.0 mg of high purity (over 98%) rhdaintain/AIF-1 was obtained from 1 L culture. The recombinant peptide was able to increase blood glucose elimination rates and enhance the proliferation of human MCF-7 cells. These results suggest that biological activity of the recombinant peptide was preserved after purification

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