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

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

Volume: 17 Issue: 2

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

Affiliation:

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

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.

Cite this article as:

Xu Zhinan, Lian Jiazhang and Cai Jin, Efficient Expression of Aquaporin Z in Escherichia coli Cell-Free System Using Different Fusion Vectors, Protein & Peptide Letters 2010; 17 (2) . https://dx.doi.org/10.2174/092986610790225996