Cry1Ab/Cry1Ah杂合蛋白构建与功能研究

doi:10.13560/j.cnki.biotech.bull.1985.2015.09.012

生物技术通报 ›› 2015, Vol. 31 ›› Issue (9): 91-96.doi: 10.13560/j.cnki.biotech.bull.1985.2015.09.012

Cry1Ab/Cry1Ah杂合蛋白构建与功能研究

徐曼¹, 蒋健², 束长龙², 张杰^1，2, 宋福平^1，2

(1东北农业大学，哈尔滨 150030；2.中国农业科学院植物保护研究所植物病虫害国家重点实验室，北京 100193)

收稿日期:2015-03-06 出版日期:2015-09-15 发布日期:2015-09-16
作者简介:徐曼，女，硕士研究生，研究方向：分子遗传学；E-mail：dianxin5@126.com
基金资助:
国家转基因专项(2014ZX0800912B，2014ZX08009003-001-004)

Creation of Cry1Ab/Cry1Ah Hybrid Proteins and Its Functional Analysis

Xu Man¹, Jiang Jian², Shu Changlong², Zhang Jie^1，2, Song Fuping^1，2

(1. Northeast Agricultural University，Harbin 150030；2. State Key Laboratory of Plant Diseases and Insect Pests，Institute of Plant Protection of Chinese Academy of Agricultural Sciences，Beijing 100193)

Received:2015-03-06 Published:2015-09-15 Online:2015-09-16

摘要/Abstract

摘要： Cry1A类杀虫蛋白是目前应用最为广泛的杀虫蛋白，目前已经报道的Cry1A类杀虫蛋白之间存在普遍的结构域交换现象。针对鳞翅目害虫具有高活性的Cry1Ab与Cry1Ah蛋白开展研究，构建了Cry1Ab、Cry1Ah的杂合蛋白AhAhAb并测定了杀虫活性。结果显示，Cry1Ab、Cry1Ah的结构域交换引起蛋白杀虫活性的显著变化，与出发蛋白相比，杂合蛋白AhAhAb丧失了对棉铃虫杀虫活性，降低了对玉米螟、小菜蛾杀虫活性。利用生物信息学方法对Cry1Ah结构域I建模，并分析其与其他Cry1A蛋白结构及表面性质差异，分析表明Cry1Ah与Cry1Ab的结构域I有相同的碳骨架和二级结构，但是表面电势分布有较大差异。进一步分析杂合蛋白AhAhAb与Cry1Ab、Cry1Ah之间杀虫活性差异的原因对进一步揭示Cry1A类蛋白杀虫特异性进化规律有重要意义。

关键词: Cry1Ah, 结构域I, 杀虫活性

Abstract: Cry1A is the most widely used insecticidal protein, and the domain swapping in the reported Cry1A family frequently occurs. In this paper, we investigated the Lepidoptera pests’ highly active protein Cry1Ab and Cry1Ah, and constructed the hybrid protein AhAhAb from Cry1Ab/Cry1Ah and bioassayed the activities. The results showed that the domain swapping of two proteins Cry1Ab and Cry1Ah resulted in the insecticidal activities significantly changed. Compared with the origin proteins, hybrid protein AhAhAb lost the insecticidal activity to Heliothis armigera, caused the insecticidal activity to Ostrinia nubilalis, Plutella xylostella reduced. We then modeled Cry1Ah domain I, and compared the structure and the surface properties with other Cry1A proteins by bioinformatics methods. The results indicated that domain I of Cry1Ah and Cry1Ab had identical carbon skeleton and secondary structure but varied surface and potential distribution. The further studies on the reasons of the activity’s difference between hybrid protein AhAhAb and Cry1Ab or Cry1Ah will give enlightenment for unclosing the evolution pattern of insecticidal specificity of Cry1A family proteins.

Key words: Cry1Ah, domain I, insecticidal activity

徐曼, 蒋健, 束长龙, 张杰, 宋福平. Cry1Ab/Cry1Ah杂合蛋白构建与功能研究[J]. 生物技术通报, 2015, 31(9): 91-96.

Xu Man, Jiang Jian, Shu Changlong, Zhang Jie, Song Fuping. Creation of Cry1Ab/Cry1Ah Hybrid Proteins and Its Functional Analysis[J]. Biotechnology Bulletin, 2015, 31(9): 91-96.

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Cry1Ab/Cry1Ah杂合蛋白构建与功能研究

Creation of Cry1Ab/Cry1Ah Hybrid Proteins and Its Functional Analysis

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