Cloning, Expression, Purification, and Insecticidal Activity of a Novel Cry1Na3 Toxin From Bacillus thuringiensis BRC-ZYR2
Cloning, Expression, Purification, and Insecticidal Activity of a Novel Cry1Na3 Toxin From Bacillus thuringiensis BRC-ZYR2
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DOI：
10.1093/jee/tow031
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发表时间：
2016-06-01
影响因子：
2.2
通讯作者：
Huang, Tianpei
中科院分区：
文献类型：
--
作者：
Dai, Ruiqing;Su, Xiaoyu;Huang, Tianpei
苏云金芽孢杆菌产生多种杀虫晶体蛋白（ICP）。基因组测序是检测和鉴定B的一种有前途的策略。苏云金杆菌ICPs，这是生物防治领域的极大兴趣。本研究从B中克隆了一个新的ICP基因。基于来自454 GS-FLX Titanium测序的基因组数据和使用B.苏云金杆菌毒素扫描仪（http：//bcam.hzaubmb.net/BtToxin_scanner/index.php）。cry 1 Na 3由B表示。苏云金杆菌毒素命名委员会（Thuringiensis Toxin Nomenclature Committee）编码的601个氨基酸，68.0-kDa蛋白与Cry 1 Na 1和Cry 1 Na 2具有95%的同源性。Cry 1 Na 3含有三个在三结构域ICP中常见的保守结构域。Cry 1 Na 3对小菜蛾Plutella xylostella（L.）和Ostriniacalis（Guenee）的LC_（50）分别为3.69 μ g/ml和31.30 μ g/ml。然而，灰飞虱striatellus（Fallen）不受影响时，喂养纯化Cry 1 Na 3（250 μ g/ml）在他们的饮食。甜菜夜蛾（Spodoptera exigua（Hubner））和鲍氏叶蝉（Colaphellus bowringi（Baly））幼虫即使在Cry 1 Na 3蛋白浓度达到500 μ g/ml时也存活。Cry 1 Na 3是一种很有前途的防治鳞翅目害虫的药剂。
Bacillus thuringiensis produces a variety of insecticidal crystal proteins (ICPs). Genome sequencing is a promising strategy for detecting and identifying B. thuringiensis ICPs, which are of great interest to the biocontrol field. In this study, a novel ICP gene was cloned from B. thuringiensis BRC-ZYR2 based on genomic data from 454 GS-FLX Titanium sequencing and an analysis of the results using the B. thuringiensis Toxin_Scanner (http://bcam.hzaubmb.net/BtToxin_scanner/index.php). cry1Na3 designated by the B. thuringiensis Toxin Nomenclature Committee, encoded a 601-amino acid, 68.0-kDa protein that exhibited 95% identity with Cry1Na1 and 99% identity with Cry1Na2. Cry1Na3 contained three conserved domains commonly found in three-domain ICPs. Cry1Na3 was toxic to Plutella xylostella (L.) and Ostrinia furnacalis (Guenee), with LC50 values of 3.69 mu g/ml and 31.30 mu g/ml, respectively. However, Laodelphax striatellus (Fallen) nymphs were unaffected when fed purified Cry1Na3 (250 mu g/ml) in their diet. Spodoptera exigua (Hubner) and Colaphellus bowringi (Baly) larvae survived even when the concentration of Cry1Na3 protein reached 500 mu g/ml. Cry1Na3 is a promising agent for the control of lepidopteran insect pests.