Journal of Integrative Agriculture ›› 2015, Vol. 14 ›› Issue (8): 1598-1603.DOI: 10.1016/S2095-3119(14)60950-3

• 论文 • 上一篇    下一篇

Comparison and optimization of the method for Cry1Ac protoxin preparation in HD73 strain

 ZHOU Zi-shan, YANG Su-juan, SHU Chang-long, SONG Fu-ping, ZHOU Xue-ping, ZHANG Jie   

  1. 1、State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310012, P.R.China
    2、State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
  • 收稿日期:2014-09-23 出版日期:2015-08-05 发布日期:2015-08-06
  • 通讯作者: ZHANG Jie, Tel: +86-10-62815921,E-mail: jzhang@ippcaas.cn
  • 作者简介:ZHOU Zi-shan, Tel: +86-10-62815612, E-mail: zishanzhou@126.com;
  • 基金资助:

    This study was supported by the National Natural Science Foundation of China (31272115), the National High-Tech R&D Program (863 Program 2011AA10A203), and the China Postdoctoral Science Foundation (2014M560487).

Comparison and optimization of the method for Cry1Ac protoxin preparation in HD73 strain

 ZHOU Zi-shan, YANG Su-juan, SHU Chang-long, SONG Fu-ping, ZHOU Xue-ping, ZHANG Jie   

  1. 1、State Key Laboratory of Rice Biology, Institute of Biotechnology, Zhejiang University, Hangzhou 310012, P.R.China
    2、State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China
  • Received:2014-09-23 Online:2015-08-05 Published:2015-08-06
  • Contact: ZHANG Jie, Tel: +86-10-62815921,E-mail: jzhang@ippcaas.cn
  • About author:ZHOU Zi-shan, Tel: +86-10-62815612, E-mail: zishanzhou@126.com;
  • Supported by:

    This study was supported by the National Natural Science Foundation of China (31272115), the National High-Tech R&D Program (863 Program 2011AA10A203), and the China Postdoctoral Science Foundation (2014M560487).

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摘要： Bacillus thuringiensis is one of the most widely used bioinsecticides, and cry gene is the major insecticidal gene. Because Cry1Ac protein shows strong toxicity against many lepidopteran species, it has been applied widely in spraying products and transgenic Bt-crops. The preparation of Cry protoxin is the first step in the very important processes of understanding the insecticidal mechanism, resistance screening, and biosafety assessments. The media for crystal production and the method for Cry protoxin preparation were varied, however, it was not clear which was better for preparing a larger amount of Cry protoxin. In this paper, three media for crystal production and the method for Cry1Ac protoxin preparation from HD73 strain were compared to find an efficacious way to prepare a large number of Cry1Ac protoxin. The results showed that the 1/2 LB (Luria-Bertani) medium was the ideal medium for crystal production, because the total yield of Cry1Ac protoxin in 300 mL 1/2 LB medium was (112.38±5.64) mg, the highest one among three media; the repeated crystal solubilization method was better for the preparation of the Cry protoxin comparing with the continuous crystal solubilization method. It will be a reference for other Cry protoxin preparation, especially for larger number.

关键词: Bacillus thuringiensis , Cry1Ac protoxin , crystal solubilization , total yield , toxicity

Abstract: Bacillus thuringiensis is one of the most widely used bioinsecticides, and cry gene is the major insecticidal gene. Because Cry1Ac protein shows strong toxicity against many lepidopteran species, it has been applied widely in spraying products and transgenic Bt-crops. The preparation of Cry protoxin is the first step in the very important processes of understanding the insecticidal mechanism, resistance screening, and biosafety assessments. The media for crystal production and the method for Cry protoxin preparation were varied, however, it was not clear which was better for preparing a larger amount of Cry protoxin. In this paper, three media for crystal production and the method for Cry1Ac protoxin preparation from HD73 strain were compared to find an efficacious way to prepare a large number of Cry1Ac protoxin. The results showed that the 1/2 LB (Luria-Bertani) medium was the ideal medium for crystal production, because the total yield of Cry1Ac protoxin in 300 mL 1/2 LB medium was (112.38±5.64) mg, the highest one among three media; the repeated crystal solubilization method was better for the preparation of the Cry protoxin comparing with the continuous crystal solubilization method. It will be a reference for other Cry protoxin preparation, especially for larger number.

Key words: Bacillus thuringiensis , Cry1Ac protoxin , crystal solubilization , total yield , toxicity