Department of Horticulture, Seoul National University, Suwon, 441-744, Korea
To facilitate construction of physical map of the hot pepper genome, a bacterial artificial chromosome (BAC) library of Capsicum annuum AC 2258, the Phytophthora capsici resistance line, is under construction. The BAC vector, pBeloBAC11, was isolated using the Plasmid Maxi kit (Qiagen) followed by cesium chloride density gradient centrifugation. The vector was digested to completion with BamHI followed by dephosphorylation with CIP. Nuclei were isolated from 60 g of young leaves using method by Zhang et al., embeded in low-melting point agarose plugs and chopped into small pieces. The maximum amount of DNA in the 200-400 kb range were used in the ligation and transformation experiments. The pepper BAC library so far, as of August 1998, consists of over 3,000 clones. The white recombinant BAC clones were characterized and stored in 384-well plates at -80”ĘC for long term storage. We are collecting BAC clones, testing the clone stability and estimating chimerism by fluorescence in situ hybridization (FISH).
Plant & Animal Genome VII Conference Plant & Animal Genome VII Conference