Cloning and expression of <Emphasis Type="Italic">Tenebrio molitor</Emphasis> antifreeze protein in <Emphasis Type="Italic">Escherichia coli</Emphasis> |
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Authors: | Chang-Wu Yue Yi-Zheng Zhang |
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Institution: | (1) College of Life Science, Sichuan Key Laboratory of Molecular Biology & Biotechnology, Sichuan University, Chengdu, 610064, P.R. China;(2) Central Laboratory of Zunyi Medical College, Zunyi, 563000, P.R. China |
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Abstract: | A novel antifreeze protein cDNA was cloned by RT-PCR from the larva of the yellow mealworm Tenebrio molitor. The coding fragment of 339 bp encodes a protein of 112 amino acid residues and was fused to the expression vectors pET32a
and pTWIN1. The resulted expression plasmids were transformed into Escherischia coli strains BL21 (DE3), ER2566, and Origami B (DE3), respectively. Several strategies were used for expression of the highly
disulfide-bonded β-helix-contained protein with the activity of antifreeze in different expression systems. A protocol for
production of refolded and active T. molitor antifreeze protein in bacteria was obtained. |
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Keywords: | Antifreeze protein Gene cloning Gene expression Inclusion body Tenebrio molitor |
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