苏云金芽孢杆菌日本血清型菌株Buibui中对金龟子有特异性的杀幼虫毒素蛋白的特性分析。
Characterization of larvicidal toxin protein from Bacillus thuringiensis serovar japonensis strain Buibui specific for scarabaeid beetles.
作者信息
Hori H, Suzuki N, Ogiwara K, Himejima M, Indrasith L S, Minami M, Asano S, Sato R, Ohba M, Iwahana H
机构信息
Tsukuba Laboratories for Research and Development, Kubota Corp., Ibaraki, Japan.
出版信息
J Appl Bacteriol. 1994 Apr;76(4):307-13. doi: 10.1111/j.1365-2672.1994.tb01633.x.
The delta-endo toxin proteins from Bacillus thuringiensis which kill the larvae of various scarabaeid beetles such as Anomala cuprea, A. rufocuprea and Popillia japonica were purified by DEAE ion exchange chromatography. A protein with a molecular size of 130 kDa was purified. During the purification a minor peak was also detected which was estimated to be 67 kDa by SDS-PAGE. Both 130 and 67 kDa proteins showed larvicidal activity against A. cuprea. The lethal concentration of the 130 kDa protein which killed 50% of the larvae tested (LC50) against A. cuprea was 2 micrograms g-1 compost. A comparison by SDS-PAGE of the V8 protease digestion pattern of the 130 and 67 kDa larvicidal proteins showed that proteolytic resistant core peptides of approximately 60 kDa molecular size were resulted. The N-terminus amino acid sequence of the 130 and 67 kDa proteins was determined to be NH2-XXPNNQNEYEIIDAL and NH2-XSRNPGTFI, respectively, which is not identical to the sequence of CryIA, CryIB, CryIC and CryIII proteins.
苏云金芽孢杆菌中能杀死多种金龟子幼虫（如铜绿异丽金龟、红脚异丽金龟和日本丽金龟）的δ-内毒素蛋白，通过DEAE离子交换色谱法进行了纯化。纯化得到了一种分子量为130 kDa的蛋白质。在纯化过程中还检测到一个小峰，通过SDS-PAGE估计其分子量为67 kDa。130 kDa和67 kDa的蛋白质对铜绿异丽金龟均表现出杀幼虫活性。130 kDa蛋白质对铜绿异丽金龟的致死浓度（杀死50%受试幼虫的LC50）为2微克/克堆肥。通过SDS-PAGE对130 kDa和67 kDa杀幼虫蛋白的V8蛋白酶消化模式进行比较，结果显示产生了分子量约为60 kDa的抗蛋白水解核心肽段。130 kDa和67 kDa蛋白质的N端氨基酸序列分别确定为NH2-XXPNNQNEYEIIDAL和NH2-XSRNPGTFI，这与CryIA、CryIB、CryIC和CryIII蛋白的序列不同。
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