Insecticidal activity of the CryIIA protein from the NRD-12 isolate of Bacillus thuringiensis subsp. kurstaki expressed in Escherichia coli and Bacillus thuringiensis and in a leaf-colonizing strain of Bacillus cereus
Insecticidal activity of the CryIIA protein from the NRD-12 isolate of Bacillus thuringiensis subsp. kurstaki expressed in Escherichia coli and Bacillus thuringiensis and in a leaf-colonizing strain of Bacillus cereus
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苏云金芽孢杆菌亚种 NRD-12 分离株 CryIIA 蛋白的杀虫活性
DOI：
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发表时间：
1994
影响因子：
4.4
通讯作者：
Paul A. Backman
中科院分区：
文献类型：
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作者：
J. William;Moar;J. Trumble;Robert H. Hice;Paul A. Backman
从苏云金芽孢杆菌亚种NRD-12分离株中扩增出一个4.0kb的BamHI-HindIII片段，编码CryIIA操纵子。将kurstaki基因克隆到大肠杆菌中。含有NRD-12基因的2.2kb ACCI-HindIII片段的核苷酸序列与HD-1和HD-263的序列完全相同。CryIIA的表达和包涵体的纯化得到了一种对绿色日光夜蛾、甜菜夜蛾和致倦库蚊具有杀虫活性的蛋白，但没有甜菜夜蛾的活性。将编码CryIIA操纵子的4.0kb BamII-HindIII片段插入苏云金杆菌E.Coli穿梭载体pHT3101(PMAU1)。用pMAU1转化苏云金芽孢杆菌HD-1无晶菌株。Kurstaki和蜡状芽孢杆菌(Bt-8)的叶部定植菌株(Bt-8)。两个转化菌株的孢子-晶体混合物对绿色隐翅虫和甜菜夜蛾均有毒力，但对玉米夜蛾和甜菜夜蛾无毒害作用。
A 4.0-kb BamHI-HindIII fragment encoding the cryIIA operon from the NRD-12 isolate of Bacillus thuringiensis subsp. kurstaki was cloned into Escherichia coli. The nucleotide sequence of the 2.2-kb AccI-HindIII fragment containing the NRD-12 cryIIA gene was identical to the HD-1 and HD-263 cryIIA gene sequences. Expression of cryIIA and subsequent purification of CryIIA inclusion bodies resulted in a protein with insecticidal activity against Heliothis virescens, Trichoplusia ni, and Culex quinquefasciatus but not Spodoptera exigua. The 4.0-kb BamII-HindIII fragment encoding the cryIIA operon was inserted into the B. thuringiensis-E. coli shuttle vector pHT3101 (pMAU1). pMAU1 was used to transform an acrystalliferous HD-1 strain of B. thuringiensis subsp. kurstaki and a leaf-colonizing strain of B. cereus (BT-8) by using electroporation. Spore-crystal mixtures from both transformed strains were toxic to H. virescens and T. ni but not Helicoverpa zea or S. exigua.
DOI：
10.1073/pnas.82.4.1074
发表时间：
1985-01-01
影响因子：
11.1
作者：
TABOR, S;RICHARDSON, CC
通讯作者：
RICHARDSON, CC
使用小量制备质粒 DNA 通过 Sequenase 对双链模板进行测序。
影响因子：
2.7
作者：
Kraft,R;Tardiff,J;Krauter,KS;Leinwand,LA
通讯作者：
Leinwand,LA