Applied Microbiology and Biotechnology, Vol.47, No.4, 379-384, 1997
Cloning and Expression of the Bacillus-Sphaericus-2362 Mosquitocidal Genes in a Nontoxic Unicellular Cyanobacterium, Synechococcus Pcc6301
Genes encoding the mosquitocidal binary toxin of Bacillus sphaericus 2362 were introduced into Synechococcus PCC6301, a cyanobacterium that can tolerate a number of potential variations in the mosquito breeding environment, and can serve as a food source for mosquito larvae. The toxin genes, preceded by a Synechococcus rbcL promoter, were located on a mobilizable Escherichia coli Synechococcus shuttle vector, which was introduced into Synechococcus PCC6301 at frequencies of 10(-5)-10(-7) exconjugants/recipient, depending on the selective conditions used. Recombinant Synechococcus exhibited significant toxicity against 2-day-old and 6-day-old Culex quinquefasciatus larvae, the concentration required to kill 50% of larvae (LC50) being 2.1 x 10(5) and 1.3 x 10(5) cells/ml respectively. Mosquitocidal activity decreased tenfold after 20 generations of non-selective growth.
Keywords:GRAM-NEGATIVE BACTERIA;ESCHERICHIA-COLI;CONJUGATIVE TRANSFER;SYNECHOCYSTIS SP;HIGH-LEVEL;PLASMIDS;DNA;TRANSFORMATION;CONSTRUCTION;VECTOR