A biolistic approach for transformation and expression of cry 1Ac gene in shoot tips of cotton (Gossypium hirsutum)

The possibility of regenerating the tetraploid cotton Gossypium hirsutum cv NH 545 plants using Shoot tips excised from 3-5 day old. seedlings was investigated. MS basal medium along with 0.1 mg/l BAP and 0.1 mg/l NAA was found better for shooting and the root initiation was obtained using 0.3 mg/l IBA. We also tried to transfer these with Cry 1Ac gene using the particle bombardment process. Shoot tips were used as explants in transformation experiment with Cry 1Ac gene and npt II as selection marker gene driven by a CaMV 35S promoter in pBIN Bt3 plasmid vector All the conditions for optimal delivery of DNA to explants were provided. Transformed putative plantlets were selected on the MS media containing growth regulators along with 75 mg/l kanamycin. The integration of the Cry 1Ac into the genomic DNA of plant was confirmed by PCR method using both npt II gene specifics. Transient expression of Cry 1Ac protein was confirmed by dipstick method.