Abstract
The immature embryos of Gossypium arboreum cv. RG8 were co-bombarded with two plasmids H1Z and pBI121 using Biolistic® PDS-1000/He Particle Delivery System. The two plasmids were coated onto tungsten particles in the ratio 1:1 at a concentration of 100ng/μl which was 10 times less than recommended in the standard protocol. A total of 1827 embryos were bombarded in 12 different experiments. The representative embryos from each bombardment experiment were analysed 48 hr post bombardment through beta glucuronidase assay called GUS assay and PCR. The histochemical results showed indigo colouration in the representative embryos from 11 experiments (91.6%). The gene specific PCR analysis on genomic DNA isolated 48 hr post bombardment from 12 representative bombarded embryos revealed 1764bp amplicon corresponding to β-1, 3-glucanase ORF in nine (75%) bombarded embryos. The results confirmed successful co-bombardment of two transgenes despite using ten times less concentration of plasmid DNA.
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