数字农科院2.0

Construction and analysis of Sip1Aa insecticidal protein random recombination library

文献类型： 外文期刊

作者： Ding Yue-ming;Wang Jing;Wang Lin;Gao Ji-Guo;Liu Rong-Mei;Li Hai-Tao

作者机构：

关键词： Bacillus thuringiensis; Colaphellus bowringi Baly; Directed evolution; Random recombination library; Sip1Aa

期刊名称： Journal of Asia-Pacific Entomology

ISSN： 1226-8615

年卷期： 2021 年 24 卷 2 期

页码：

收录情况： JCR（2021版）

摘要： The Sip1Aa protein from Bacillus thuringiensis is highly toxic to Colaphellus bowringi Baly. In order to obtain mutant proteins with higher insecticidal activity, a random recombinant library of Sip1Aa protein was constructed using error-prone PCR. A total number of 100 positive transformants were randomly selected for sequence determination, and 25 mutants (M1 to M25) were selected and expressed the respective Sip1Aa mutants. These Sip1Aa variants had a total of 29 base mutations, with an average of 1.2 base mutations per mutant. Compared with that of the wild-type Sip1Aa protein, the insecticidal activity of the mutants M1 (A31G, Y118C, D227E), M5 (K168R) and M21 (I307T) was significantly decreased, with and LC50 values 4 to 6 times higher than the Sip1Aa protein. The mutant M8 (R174S) showed increase in the insecticidal activity against the Colaphellus bowringi Baly was obtained, with an LC50 value 4-fold less than the Sip1Aa protein. The results of this study provide reference for the molecular modification of Sip1Aa protein and the study of key sites of its insecticidal activity. © 2021 Korean Society of Applied Entomology

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