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蜡样芽孢杆菌群菌株中编码cry1Ac基因的质粒的接合转移、稳定性及表达

Conjugative transfer, stability and expression of a plasmid encoding a cry1Ac gene in Bacillus cereus group strains.

作者信息

Hu Xiaomin, Hansen Bjarne Munk, Eilenberg Jørgen, Hendriksen Niels Bohse, Smidt Lasse, Yuan Zhiming, Jensen Gert Bolander

机构信息

National Environmental Research Institute, 4000 Roskilde, Denmark.

出版信息

FEMS Microbiol Lett. 2004 Feb 9;231(1):45-52. doi: 10.1016/S0378-1097(03)00925-X.

Abstract

The plasmid pHT73 containing cry1Ac and tagged with an erythromycin resistance gene was transferred from Bacillus thuringiensis subspecies kurstaki KT0 to several Bacillus cereus group strains by conjugation. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and phase contrast microscopy showed that the transconjugants containing plasmid pHT73 could express Cry1Ac toxin and produce bipyramidal crystalline inclusion bodies during sporulation. The study demonstrated that pHT73 could be transferred to B. thuringiensis subsp. kurstaki, several B. cereus strains and Bacillus mycoides. Under non-selective conditions, the stability of the pHT73 plasmid in the transconjugants was found to be 58.2-100% after 100 generations and 4-96% after 200 generations. The variations are mainly caused by the choice of receptor strain.

摘要

携带cry1Ac并标记有红霉素抗性基因的质粒pHT73通过接合作用从苏云金芽孢杆菌库斯塔克亚种KT0转移至几种蜡样芽孢杆菌属菌株。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和相差显微镜观察表明,含有质粒pHT73的接合子在孢子形成过程中能够表达Cry1Ac毒素并产生双锥体晶体包涵体。该研究证明pHT73能够转移至苏云金芽孢杆菌库斯塔克亚种、几种蜡样芽孢杆菌菌株和蕈状芽孢杆菌。在非选择性条件下,发现接合子中pHT73质粒在传代100次后的稳定性为58.2%-100%,传代200次后的稳定性为4%-96%。这些变化主要是由受体菌株的选择引起的。

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