Construction and analysis of Sip1Aa insecticidal protein random recombination library
文献类型: 外文期刊
第一作者: Ding, Yue-ming
作者: Ding, Yue-ming;Wang, Jing;Wang, Lin;Gao, Ji-Guo;Liu, Rong-Mei;Li, Hai-Tao;Li, Hai-Tao
作者机构:
关键词: Bacillus thuringiensis; Sip1Aa; Random recombination library; Directed evolution; Colaphellus bowringi Baly
期刊名称:JOURNAL OF ASIA-PACIFIC ENTOMOLOGY ( 影响因子:1.303; 五年影响因子:1.427 )
ISSN: 1226-8615
年卷期: 2021 年 24 卷 2 期
页码:
收录情况: SCI
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.
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