Improving the insecticidal activity by expression of a recombinant cry1Ac gene with chitinase-encoding gene in acrystalliferous Bacillus thuringiensis
Improving the insecticidal activity by expression of a recombinant cry1Ac gene with chitinase-encoding gene in acrystalliferous Bacillus thuringiensis
复制标题
含几丁质酶编码基因的重组cry1Ac基因在晶体苏云金芽孢杆菌中表达提高杀虫活性
DOI：
10.1007/s00284-008-9112-1
复制
发表时间：
2008-05-01
影响因子：
2.6
通讯作者：
Zhang, Youming
中科院分区：
文献类型：
--
作者：
Ding, Xuezhi;Luo, Zhaohui;Zhang, Youming
为了提高几丁质酶的杀虫活性，将烟草几丁质酶基因和苏云金芽孢杆菌cry 1Ac基因分别克隆到pHT 315载体上，构建成pHUAccB 5质粒。在天然cry 1Ac启动子的控制下尝试构建转录融合体。将质粒pHUAccB 5导入B。苏云金杆菌无晶体的电穿孔。SDS-PAGE和Western blot分析表明，该菌株能表达130 kDa的Cry 1Ac蛋白和30 kDa的几丁质酶蛋白。在几丁质酶活性分析中，菌株XBU-HUAccB 5的几丁质酶活性在72 h时达到7.5 U/mL，是HTX-42的5倍，是亲本菌株的6倍。以棉铃虫Helicoverpa armigera Hubner为试虫，测定了XBU-HUAccB 5的杀虫活性，在48 h时，XBU-HUAccB 5的杀虫活性是表达cry 1Ac的HTX-42的11.30倍，在72 h时，XBU-HUAccB 5的杀虫活性是表达cry 1Ac的HTX-42的18.76倍。
In order to improve the insecticidal activity, the chitinase gene from tobacco (Nicotiana tabacum) endochitinase and the cry1Ac gene from Bacillus thuringiensis were cloned into the vector pHT315 and designated as pHUAccB5 plasmid. The constructed transcriptional fusion was attempted under the control of the native cry1Ac promoter. Plasmid pHUAccB5 was introduced into B. thuringiensis acrystalliferous by electroporation. Analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot, the transformant XBU-HUAccB5 produced 130-kDa Cry1Ac protein and 30-kDa chitinase protein. During the chitinase active analysis, the transformant, XBU-HUAccB5 chitinase active, reached 7.5 U/mL at 72 h, and was 5 times higher than the HTX-42 and 6 times higher than the parent strains. When the insecticidal activity of the transformant was evaluated against Helicoverpa armigera Hubner, the XBU-HUAccB5 toxicity was 11.30 times higher than the transformant HTX-42 expressed single cry1Ac at 48 h and was 18.76 times higher at 72 h.