Efficient Expression of Aquaporin Z in Escherichia coli Cell-Free System Using Different Fusion Vectors

Author(s): Zhinan Xu, Jiazhang Lian, Jin Cai.

Journal Name: Protein & Peptide Letters

Volume 17 , Issue 2 , 2010

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Abstract:

Aquaporin Z (AqpZ) is a typical orthodox aquaporin with 6 transmembrane domains and five connecting loops. In order to express this complex membrane protein efficiently, E. coli cell-free expression system was employed as an alternative to produce aquaporin Z. Using different fusion vectors containing AqpZ gene, the expression level of fusion proteins in cell-free system varied from 7.97 to 578.35 μg/ml, while 7.34 to 340.81 μg/ml for target protein (AqpZ). The free energy of mRNA secondary structure at translation initiation region (TIR) was predicted and demonstrated a positive relationship with the expression level of AqpZ in cell-free system. This is the first report of expressing water channel protein in E. coli cell-free system, which has become a highly promising tool for fast and efficient production of integral membrane proteins.

Keywords: Aquaporin, Aquaporin Z, Cell-free expression system, mRNA secondary structure, Membrane protein

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Article Details

VOLUME: 17
ISSUE: 2
Year: 2010
Page: [181 - 185]
Pages: 5
DOI: 10.2174/092986610790225996
Price: $58

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