In vivo fluorescence observation of parasporal inclusion formation in Bacillus thuringiensis

文献类型： 外文期刊

第一作者： Rong Rong

作者： Rong Rong;Song FuPing;Sun ChangPo;Wei Juan;Zhang Jie;Yang Hui;Huang DaFang

作者机构：

关键词： Bacillus thuringiensis;CrylAc-GFP fusion protein;laser confocal microscopy

期刊名称：SCIENCE CHINA-LIFE SCIENCES （ 影响因子：6.038； 五年影响因子：4.754 ）

ISSN： 1674-7305

年卷期： 2010 年 53 卷 9 期

页码：

收录情况： SCI

摘要： A recombinant gene expressing a CrylAc-GFP fusion protein with a molecular mass of approximately 160 kD was constructed to investigate the expression of crylAc, the localization of its gene product CrylAc, and its role in crystal development in Bacillus thuringiensis. The crylAc-gfp fusion gene under the control of the crylAc promoter was cloned into the plasmid pHT304, and this construct was designated pHTcrylAc-gfp. pHTcrylAc-gfp was transformed into the crystal-negative strain, HD-73 cry(-), and the resulting strain was named HD-73(-)(pHTcrylAc-gfp). The gfp gene was then inserted into the large HD-73 endogenous plasmid pHT73 and fused with the 3' terminal of the ctylAc gene by homologous recombination, yielding HD-730 Phi(crylAc-gfp)3534. Laser confocal microscopy and Western blot analyses showed for the first time that the CrylAc-GFP fusion proteins in both HD-73(-)(pHTcrylAc-gfp) and HD-730 Phi(crylAc-gfp)3534 were produced during asymmetric septum formation. Surprisingly, the CrylAc-GFP fusion protein showed polarity and was located near the septa in both strains. There was no significant difference between CrylAc-GFP and CrylAc in their toxicity to Plutella xylostella larvae.

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