Isolation, characterization and expression of a novel vegetative insecticidal protein gene of Bacillus thuringiensis
Isolation, characterization and expression of a novel vegetative insecticidal protein gene of Bacillus thuringiensis
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DOI：
10.1016/j.femsle.2005.01.011
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发表时间：
2005-02-15
影响因子：
2.1
通讯作者：
Kumar, PA
中科院分区：
文献类型：
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作者：
Bhalla, R;Dalal, M;Kumar, PA
利用vip 3Aa 1特异性引物，对24个苏云金芽孢杆菌（Bt）血清型进行了vip样基因的PCR检测。在8个血清型中鉴定VIP样基因序列。这些基因被克隆和测序。苏云金芽孢杆菌tolivipAa14基因推导的氨基酸序列与目前报道的Vips有很大的差异。利用表达载体pET 29a在大肠杆菌中表达了Bt tolwarthi vip3Aa14基因。表达的Vip3Aa14蛋白在胞质上清液以及沉淀级分中被发现，但蛋白质在胞质上清液级分中更丰富。全长和截短的Vips对斜纹夜蛾和小菜蛾的幼虫都有很高的毒力。Vip3Aa14蛋白N端截短不影响其杀虫活性。(C)2005年，欧洲微生物学会联合会。Elsevier B.V.出版，保留所有权利。
Twenty-four serovars of Bacillus thuringiensis (Bt) were screened by polymerase chain reaction to detect the presence of vegetative insecticidal protein gene (vip)-like sequences by using vip3Aa 1-specific primers. vip-like gene sequences were identified in eight serovars. These genes were cloned and sequenced. The deduced amino acid sequence of the vipAa14 gene from Bacillus thuringiensis tolworthi showed considerable differences as compared to those of Vips reported so far. The vip3Aa14 gene from Bt tolwarthi was expressed in Eseherichia coli using expression vector pET29a. The expressed Vip3Aa14 protein was found in cytosolic supernatant as well as pellet fraction, but the protein was more abundant in the cytosolic supernatant fraction. Both full-length and truncated (devoid of signal sequence) Vips were highly toxic to the larvae of Spodoptcra litura and Plutella xylostella. Truncation of Vip3Aa14 protein at N-terminus did not affect its insecticidal activity. (C) 2005 Federation of European Microbiological Societies. Published by Elsevier B.V. All rights reserved.