Effect of Regular Repetitive Sequence on the Thermal Hysteresis Activity of Antifreeze Protein ApAFP914 in Anatolica polita

doi:10.13560/j.cnki.biotech.bull.1985.2017.02.027

Abstract

Abstract: This work aims to explore the relationship between protein structure and thermal hysteresis activity of antifreeze protein ApAFP914 in Anatolica polita. A regular repetitive sequence was added in the ApAFP914 by gene synthesis. The synthesized gene was then expressed in prokaryotic cells and the thermal hysteresis activity of the protein was determined. Results showed that the gene ApAFP-C914 with a regular repetitive sequence added was 312 bp. The fusion protein TrxA-ApAFP-C914 was verified by SDS-PAGE and Western bolt,and its molecular weight was 34 kD. The measurement by differential scanning calorimetry showed that the thermal hysteresis activity of ApAFP increased significantly（P<0.05）by adding a regular repetitive sequence at the concentration of 50 μg/mL. Conclusively,adding repetitive sequence may significantly increase the thermal hysteresis activity of antifreeze protein ApAFP914 in A. polita.

Key words: antifreeze protein, differential scanning calorimetry, repetitive sequence, thermal hysteresis

WANG Jun, DU Rong-feng, LIU Zhong-yuan, MAO Xin-fang. Effect of Regular Repetitive Sequence on the Thermal Hysteresis Activity of Antifreeze Protein ApAFP914 in Anatolica polita[J]. Biotechnology Bulletin, 2017, 33(2): 185-191.

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