Cloning, Expression and Toxicity of a Mosquitocidal Toxin Gene of Bacillus thuringiensis subsp. medellin

Mosquitocidal bacterial toxins: diversity, mode of action and resistance phenomena

“…Inclusions of Btmed are composed of four different and major proteins of 94, 70-68, 30 and 28kDa (Restrepo et al 1997). The 94kDa protein, named Cry11Bb (as it is 83% identical to Cry11Ba) is the most toxic component, whatever the mosquito species is, with a level of activity comparable to that of Cry11Ba (Orduz et al 1998).…”

Expression of mosquito active toxin genes by a Colombian native strain of the gram-negative bacterium Asticcacaulis excentricus

“…B. sphaericus strain 2362 was grown in NYSM (g/l: nutrient broth, 28; yeast extract, 0.5 and supplemented with salt solution as follows: 0.7 mM CaCl 2 , 1 mM MgCl 2 , and 50 µM MnCl 2 ) (Myers & Yousten 1978). Recombinant B. thuringiensis expressing the Cry11Bb protein was developed by Restrepo et al (1997) and cultured by using M1 liquid medium. The Cry11Bb crystals were purified from a final whole culture (FWC) as described by Pendleton and Morrison (1966) with modifications introduced by Otieno-Ayayo et al (1993).…”

“…Plasmid pBTM4 containing the cry11Bb1 gene was constructed by Restrepo et al (1997). Plasmid pEA1 containing B. sphaericus binary toxin gene was a gift from Dr Alan Porter (Institute of Molecular and Cellular Biology, National University of Singapore).…”

“…B. thuringiensis subsp. medellin was grown in M1 at 30 o C, 200 rpm, for 48 h until sporulation (Restrepo et al 1997). Ae.…”

Proteolytic processing of the Cyt1Ab1 toxin produced by Bacillus thuringiensis subsp. medellin

“…Crystal purification -Recombinant strains containing the cyt1Aa1 and cyt1Ab1 genes were incubated in 5 ml of M-one liquid medium (Restrepo et al 1997) supplemented with erythromycin (25 µg/ml), for 12 h at 30°C and 200 rpm. Cultures were transferred to 200 ml of M-one liquid medium and incubated for 120 h at 30°C and 200 rpm until sporulation.…”