In vivo fluorescence observation of parasporal inclusion formation in Bacillus thuringiensis  

作　　者：YANG Hui RONG Rong SONG FuPing SUN ChangPo WEI Juan ZHANG Jie HUANG DaFang 

机构地区：[1]State Key Laboratory for Biology of Plant Diseases and Insect Pests,Institute of Plant Protection,Chinese Academy of Agricultural Sciences,Beijing 100193,China [2]Biotechnology Research Institute,Chinese Academy of Agricultural Sciences,Beijing 100081,China

出　　处：《Science China(Life Sciences)》2010年第9期1106-1111,共6页中国科学(生命科学英文版)

基　　金：supported by the National Basic Research Program of China (Grant No. 2009CB118902);the National High-Tech Research and Development Program of China (Grant No. 2006AA10A212)

摘　　要：A recombinant gene expressing a Cry1Ac-GFP fusion protein with a molecular mass of approximately 160 kD was constructed to investigate the expression of cry1Ac,the localization of its gene product Cry1Ac,and its role in crystal development in Bacillus thuringiensis.The cry1Ac-gfp fusion gene under the control of the cry1Ac promoter was cloned into the plasmid pHT304,and this construct was designated pHTcry1Ac-gfp.pHTcry1Ac-gfp was transformed into the crystal-negative strain,HD-73 cry-,and the resulting strain was named HD-73-(pHTcry1Ac-gfp).The gfp gene was then inserted into the large HD-73 endogenous plasmid pHT73 and fused with the 3' terminal of the cry1Ac gene by homologous recombination,yielding HD-73Φ(cry1Ac-gfp)3534.Laser confocal microscopy and Western blot analyses showed for the first time that the Cry1Ac-GFP fusion proteins in both HD-73-(pHTcry1Ac-gfp) and HD-73Φ(cry1Ac-gfp)3534 were produced during asymmetric septum formation.Surprisingly,the Cry1Ac-GFP fusion protein showed polarity and was located near the septa in both strains.There was no significant difference between Cry1Ac-GFP and Cry1Ac in their toxicity to Plutella xylostella larvae.A recombinant gene expressing a Cry1Ac-GFP fusion protein with a molecular mass of approximately 160 kD was constructed to investigate the expression of cry1Ac,the localization of its gene product Cry1Ac,and its role in crystal development in Bacillus thuringiensis.The cry1Ac-gfp fusion gene under the control of the cry1Ac promoter was cloned into the plasmid pHT304,and this construct was designated pHTcry1Ac-gfp.pHTcry1Ac-gfp was transformed into the crystal-negative strain,HD-73 cry-,and the resulting strain was named HD-73-(pHTcry1Ac-gfp).The gfp gene was then inserted into the large HD-73 endogenous plasmid pHT73 and fused with the 3’ terminal of the cry1Ac gene by homologous recombination,yielding HD-73Φ(cry1Ac-gfp)3534.Laser confocal microscopy and Western blot analyses showed for the first time that the Cry1Ac-GFP fusion proteins in both HD-73-(pHTcry1Ac-gfp) and HD-73Φ(cry1Ac-gfp)3534 were produced during asymmetric septum formation.Surprisingly,the Cry1Ac-GFP fusion protein showed polarity and was located near the septa in both strains.There was no significant difference between Cry1Ac-GFP and Cry1Ac in their toxicity to Plutella xylostella larvae.

关 键 词：Bacillus thuringiensis Cry1Ac-GFP fusion protein laser confocal microscopy 

分 类 号：Q93[生物学—微生物学]