增加规则重复序列对光滑鳖甲抗冻蛋白ApAFP914热滞活性的影响

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

生物技术通报 ›› 2017, Vol. 33 ›› Issue (2): 185-191.doi: 10.13560/j.cnki.biotech.bull.1985.2017.02.027

增加规则重复序列对光滑鳖甲抗冻蛋白ApAFP914热滞活性的影响

王军, 杜荣俸, 刘忠渊, 毛新芳

新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐 830046

收稿日期:2016-05-19 出版日期:2017-02-26 发布日期:2017-02-08
作者简介:王军,男,硕士研究生,研究方向：生物化学与分子生物学;E-mail：496269587@qq.com
基金资助:
新疆维吾尔自治区自然科学基金项目（2012211A022）

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

WANG Jun, DU Rong-feng, LIU Zhong-yuan, MAO Xin-fang

Xinjiang Key Laboratory of Biological Resources and Genetic Engineering,College of Life Science and Technology,Xinjiang University,Urumqi 830046

Received:2016-05-19 Published:2017-02-26 Online:2017-02-08

摘要/Abstract

摘要： 旨在探究光滑鳖甲抗冻蛋白ApAFP914结构与热滞活性（Thermal hysteresis activity,THA）的关系。采用基因合成的方法,在光滑鳖甲抗冻蛋白基因ApAFP914中增加单个规则重复序列并进行蛋白原核表达及热滞活性测定。结果显示,增加单个重复序列的ApAFP-C914为312 bp,融合蛋白TrxA-ApAFP-C914经SDS-PAGE及Western bolt分析表明,分子量为34 kD。差示扫描量热法（Differential scanning calorimetry,DSC）测定表明,在50 μg/mL的浓度下,增加单个重复序列显著提高了ApAFP的THA活性。光滑鳖甲抗冻蛋白ApAFP914增加一个重复序列可显著提高其热滞活性。

关键词: 抗冻蛋白, 差式扫描量热法, 重复序列, 热滞活性

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

王军, 杜荣俸, 刘忠渊, 毛新芳. 增加规则重复序列对光滑鳖甲抗冻蛋白ApAFP914热滞活性的影响[J]. 生物技术通报, 2017, 33(2): 185-191.

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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增加规则重复序列对光滑鳖甲抗冻蛋白ApAFP914热滞活性的影响

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

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