Figure 6
An analysis of Hps genomic clones. A, Three independent clones isolated from soybean (Glycine max cv. Harosoy 63) genomic libraries are illustrated, along with their GenBank accession numbers. Shown for each clone are: blue box, location of the Hps gene; arrow, direction of transcription; Bm, Bg, E, and H, restriction enzyme sites for BamHI, BglII, EcoRI, and HindIII, respectively; MAR, predicted matrix attachment regions; * (asterik), location of micro-satellite dinucleotide AT repeats; Probe I, Probe II, and Probe III, correspond to DNA sequence fragments used to probe genomic DNA blots shown in the lower part of the figure. The three clones are aligned with overlapping regions of nearly identical DNA sequence. The predicted restriction enzyme fragments and their corresponding size (kb) are shown in colour directly below the genomic clones. B, Soybean (Glycine max cv. Harosoy 63) genomic DNA blot hybridizations using three different DNA probes, Probe I, Probe II, and Probe III. Samples of 20 μg of soybean genomic DNA were digested with the indicated restriction enzymes. A 5 ng sample of a plasmid (pHPS) carrying a genomic clone encoding Hps (GenBank accession number DQ208939) was digested with EcoRI, and included as an internal hybridization standard. The DNA samples were separated by agarose gel electrophoresis and blotted to Nylon membrane, prior to hybridization to each of 32P labelled DNA probes (with stripping between hybridizations). Coloured boxes and sizes (kb) of the most intense hybridizing fragments are indicated, and were matched to the predicted restriction enzyme fragments from the alignment of the genomic clones, shown in the upper part of the figure. The migration of DNA markers and their size (kb) are shown on the left.