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Journal of Integrative Agriculture  2014, Vol. 13 Issue (5): 1036-1042    DOI: 10.1016/S2095-3119(13)60532-8
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The Minimal Active Fragment of the Cry1Ai Toxin is Located Between 36I and 605I
 ZHOU Zi-shan, LIN Hui-yan, LI Ying, SHU Chang-long, SONG Fu-ping , ZHANG Jie
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
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摘要  The novel cry1Ai gene that cloned from Bacillus thuringiensis strain SC6H8 encoded a protein exhibiting strong toxicity against Plutella xylostella and Chilo suppressalis in our previous study. Using the available information for the active fragments of other Cry toxins, eight truncated fragments were constructed to identify the minimal active fragment of Cry1Ai. All truncated fragments were expressed in Escherichia coli strain BL21 (DE3), and the insecticidal activity against 2nd- instar P. xylostella larvae was assessed using full-length Cry1Ai as a positive control. The results indicate that the minimal active fragment of the Cry1Ai toxin against P. xylostella is located between amino acid residues 36I and 605I, which is smaller than the regions previously reported for Cry1A. The first two amino acids (34T and 35P) on helix α-1 and whole helix α-2 of domain I and sheet β-32 of domain III are necessary for Cry1Ai toxin to keep its toxicity against P. xylostella.

Abstract  The novel cry1Ai gene that cloned from Bacillus thuringiensis strain SC6H8 encoded a protein exhibiting strong toxicity against Plutella xylostella and Chilo suppressalis in our previous study. Using the available information for the active fragments of other Cry toxins, eight truncated fragments were constructed to identify the minimal active fragment of Cry1Ai. All truncated fragments were expressed in Escherichia coli strain BL21 (DE3), and the insecticidal activity against 2nd- instar P. xylostella larvae was assessed using full-length Cry1Ai as a positive control. The results indicate that the minimal active fragment of the Cry1Ai toxin against P. xylostella is located between amino acid residues 36I and 605I, which is smaller than the regions previously reported for Cry1A. The first two amino acids (34T and 35P) on helix α-1 and whole helix α-2 of domain I and sheet β-32 of domain III are necessary for Cry1Ai toxin to keep its toxicity against P. xylostella.
Keywords:  Bacillus thuringiensis       Cry1Ai toxin       active fragment       toxicity       Plutella xylostella  
Received: 20 March 2013   Accepted:
Fund: 

This study was supported by the National Natural ScienceFoundation of China (31272115), the National 973 Program of China (2009CB118902). We thank Ms. Liang Yingping and Liu Shuqin from Institution of Plant Protection, Chinese Academy of Agricultural Sciences, for their assistance with insect rearing and the bioassays.

Corresponding Authors:  ZHANG Jie, Tel: +86-10-62815921, Fax: +86-10-62812642, E-mail: jzhang@ippcaas.cn     E-mail:  jzhang@ippcaas.cn

Cite this article: 

ZHOU Zi-shan, LIN Hui-yan, LI Ying, SHU Chang-long, SONG Fu-ping , ZHANG Jie. 2014. The Minimal Active Fragment of the Cry1Ai Toxin is Located Between 36I and 605I. Journal of Integrative Agriculture, 13(5): 1036-1042.

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