Transformation studies of Bacillus thuringiensis cryIC gene into a nitrogen-fixing Azospirillum lipoferum.
A lepidopteran toxin gene, cryIC (pSB607) from entomopathogenic Bacillus thuringiensis subsp. aizawai was introduced into nitrogen-fixing Azospirillum lipoferum by transformation. Regeneration of spheroplasts was achieved at 99% with 39% frequency of regeneration. Transformants were screened on NB kanamycin with ampicillin plates and 4 transformants were selected after ten generations. SDS-PAGE and Western blot analysis confirmed the presence of a 68 kDa protein in the transformants. Studies on utilization of carbon sources indicate that glucose and sucrose are the most favorable carbon sources and 2% molasses is the cheap alternate carbon source for the better growth of parent A. lipoferum and transformants.
Gounder
R
R
Centre for Biotechnology, Anna University, Chennai, India.
Rajendran
N
N
D016949:Azospirillum / Q000201:enzymology* / Q000254:growth & development; D001413:Bacillus thuringiensis / Q000235:genetics* / Q000254:growth & development; D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins / Q000235:genetics*; D001427:Bacterial Toxins / Q000235:genetics; D003470:Culture Media; D004731:Endotoxins / Q000235:genetics*; D005947:Glucose / Q000378:metabolism; D006460:Hemolysin Proteins; D007700:Kinetics; D007785:Lactose / Q000378:metabolism; D008320:Maltose / Q000378:metabolism; D008353:Mannitol / Q000378:metabolism; D009586:Nitrogen Fixation; D013104:Spheroplasts / Q000502:physiology; D013395:Sucrose / Q000378:metabolism; D014169:Transformation, Bacterial
D000083722:Bacillus thuringiensis Toxins; D001426:Bacterial Proteins; D001427:Bacterial Toxins; D003470:Culture Media; D004731:Endotoxins; D006460:Hemolysin Proteins; C030956:insecticidal crystal protein, Bacillus Thuringiensis; D008353:Mannitol; D013395:Sucrose; D008320:Maltose; D005947:Glucose; D007785:Lactose