New Vector pBUC3 can Express Bt Cry1Ac31 during the Vegetative Phase in Bacillus thuringiensis
1. Hainan Institute of Tropical Agricultural Resources, Sanya, 572025, P.R. China
2. College of Life and Technology Science, Guangxi University, Nanning, 530004, P.R. China
Bt Research, 2011, Vol. 2, No. 2 doi: 10.5376/bt.2011.02.0002
Received: 08 Jul., 2011 Accepted: 12 Nov., 2011 Published: 25 Nov., 2011
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Preferred citation for this article:
Liu et al., 2011, New Vector pBUC3 can Express Bt Cry1Ac31 during the Vegetative Phase in Bacillus thuringiensis, Bt Research, Vol.2, No.2 9-13 (doi: 10.5376/bt.2011.02.0002)
The feature of Bacillus thuringiensis (B. t.) is the ability to generate parasporal crystal protein during the spore-forming process. In order to realize the expression of the Bt Cry genes independent on spore formation, we attempted to design new vector that could express Bacillus thuringiensis insecticidal crystal protein in the vegetable growth phase in this research. First, we designed a pair of specific primers based on cry3Aa promoter gene sequence to amplify cry3Aa promoter from B.t. subsp. Tenebrions; then the DNA fragment with blunt-end was ligated to the cross-species vector pBU4 to build new pBUC3 vector which would be non-dependent sporulation vector expressing in the vegetative growth phase of Bacillus thuringiensis. In order to validate the vector whether express B.t cry gene in the vegetative phase, The B.t cry1Ac31 gene was ligated to the pBUC3 vector to be transformed into B.t non crystal mutant IPS strain by electronic transformation approach. Through the examinations by the PCR, microscopic observation and SDS-PAGE, the results showed that B.t cry1Ac31 was expressed in cell growth vegetable phase. Our study suggested that the feature of cry3Aa promoter being non-dependent on spore formation might be useful expression element that could achieve the cry gene of Bacillus thuringiensis expressing in vegetative phase.
Bacillus thuringiensis; Parasporal crystal protein; cry3Aa promoter; cry1Ac31; Vegetative expression vector