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Analysis of Newly Introduced Centromeric DNA and a Linked Marker

The long‐term goal of this study is to begin developing the technology for making artificial chromosomes in corn. A BAC, bacterial artificial chromosome, containing centromeric DNA (16H10‐1), along with a plasmid containing the GUS and Basta R gene, was introduced to the end of chromosome 2. The intent was to use GUS as a visual marker for centromere activity. However, although GUS and the centromeric BAC were known to roughly co‐segregate, no direct test of linkage had been carried out. To this end, plants from three different test crosses were analyzed. If the markers are indeed tightly linked (perhaps intermingled on the chromosome), we expected all plants that are GUS‐positive to be positive for the introduced centromeric DNA. A PCR marker for the internal segment of the BAC was used to monitor its presence.