[Expression and synergism of two cry insecticidal protein genes in Pseudomonas fluorescens]

Several engineered Pseudomonas fluorescens(Pf) strains were constructed mainly based on a Pseudomonas plasmid pJMS6 alpha-lac and two insecticidal crystal protein genes of Bacillus thuringiensis, cry1Ac and cry2Aa, and the host Pf strain, P303, which was with highly antifungal activity to some plant disease fungi and colonizing ability on a wide range of plants. The DNA introduction was confirmed by PCR-RFLP and Southern blot. The 132 kD insecticidal protein was detected in IPP101 and IPP202 by SDS-PAGE and rhombic insecticidal protein crystals of them were observed through electron microscope, also indicating that cry1Ac gene was highly expressed. The results of insecticidal bioassay indicated that IPP101 was more toxic than IPP201, and IPP202 was the most toxic among the 3 strains. LC50 to the neonates of cotton boll worm(Helicoverpa armigera) were 0.02604, 0.00812 and 0.00186 mL/g feed, consecutively. In IPP202, two gene products showed significant synergism, with the co-toxicity coefficient of 332.8.