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Figure 2 | Biotechnology for Biofuels

Figure 2

From: Production of marker-free transgenic Jatropha curcas expressing hybrid Bacillus thuringiensis δ-endotoxin Cry1Ab/1Ac for resistance to larvae of tortrix moth (Archips micaceanus)

Figure 2

Southern blot analysis of transgenic Cry1Ab/1Ac plants of J. curcas. (A) Detection of T-DNA copy number in transgenic T0 plant L10 (L10-T0). Genomic DNA samples of L10-T0 and cultivar MD44 were digested with restriction enzyme NcoI. The southern blot was hybridized with a DNA probe derived from the Cry1Ab/1Ac gene. Arrow indicates the T-DNA copy that was putative marker-free (see below), which was selected for further study. (B) Transmission of the marker-free T-DNA in the T1 progeny of L10. Southern blot analysis was carried out using the restriction enzyme NcoI and the Cry1Ab/1Ac probe. L10-T1-10 and L10-T1-18 are two individual T1 plants of L10. (C) Detection of Hpt gene in T1 plants L10-T1-10 and L10-T1-18 by southern blot analysis. The same southern blot in (B) was stripped and then re-hybridized with an Hpt gene probe. (D) Detection of the intact P Pepc :Cry1Ab/1Ac:T Nos gene in the marker-free T1 progeny of L10. Genomic DNA samples of L10-T1-10, L10-T1-18 and MD44 were digested with restriction enzymes KpnI and SphI. The southern blot was hybridized with the Cry1Ab/1Ac probe. The expected size of the KpnI-SphI fragment of the P Pepc :Cry1Ab/1Ac:T Nos gene is 3452 bp.

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