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棉 花 学 报 Cotton Science 2008，20(1)：9-13
Molecular Characterization on Offsprings from a Multiple-sites Integration Transgenic Cotton （Gossypium hisutum L.） Abstract：To investigate the pattern and organization of foreign gene integration in transgenic cotton, PCR and Southern blot analysis on T0 plant，and the corresponding T1 population of a transgenic cotton ( Gossypium hirsutum L.) with multiple-integration sites derived from a simple construct pPZP-GFP via Agrobacterium -mediated transformation, and the corresponding T1 population were carried out in this paper. The seeds harvasted from GFP47 T0 and genomic DNA isolated from T0 plants were introduced from Plant Biotechnology lab of Temasek Life Sciences Laboratory，National University of Singapore. T1 plants were planted in the greenhouse for DNA isolation and thereafter molecular analysis. PCR results showed that the T0 plant was integrated not only with normal T-DNA including target gene ( gfp ) and selection marker gene ( nptII), but also with vector backbone fragment outside the left border of the T-DNA. Besides confirmed the PCR results, Southern blot analysis on T0 plant indicated that the T0 plant was integrated with multi-copy of T-DNA. PCR characterization on 33 T1 individuals showed that some individuals with only normal T-DNA structures were segregated from the population. Further analysis with Southern blot on these individuals with different genotype identified by PCR showed that there were，at least，4 loci with different integration patterns in the cotton genome. The desirable transgenic event with only single copy of normal T-DNA integration could be segregated from the offsprings of this multiple-sites integration transgenic cotton.
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