Promoter fusion with Green Fluorescent Protein (GFP) is often used to determine a gene’s transcriptional activity in vivo. To transform Arabidopsis plant with promoter-GFP transgenes, Agrobacterium inserts a transfer DNA (T-DNA) containing promoter-GFP and a kanamycin-resistant gene into the plant genome. T-DNA insertion can happen at a single locus or multiple loci. The identification of a single insertion line is necessary because it simplifies downstream genetic analysis. A transgenic line is likely a single insertion line if 75% of the progenies from a heterozygous plant are kanamycin-resistant. The goal of this study is to identify single insertion lines for InvINH1 and InvINH2 promoter-GFP fusions. For each transgenic line, 300 T2 seeds were sterilized and plated on MS medium containing 35ug/ml kanamycin. The number of kanamycin-resistant seedlings was determined two weeks after germination. In total, 11 lines were analyzed for both transgenes. For each transgene, at least two single insertion lines had been identified, and their GFP expression pattern will later be validated. In addition, these lines will be used to determine whether both the paternal and maternal InvINHs alleles are expressed in the endosperm. Confirmed lines will be deposited into a stock center and made available to the scientific community.
