Plant & Animal Genome IX Conference

Poster: Large Insert Libraries, Gene Isolation, Etc.
P02_05.html

CONSTRUCTION OF A SWEETPOTATO BAC LIBRARY AND ITS APPLICATION FOR RESISTANCE GENE ISOLATION

¹ CALS Genome Research Laboratory, North Carolina State University, Campus Box 8619, Raleigh, NC, 27695

² Department of Horticulture, North Carolina State University, Campus Box 7609, Raleigh, NC, 27695

Sweetpotato is the seventh most important crop in the world, and ranks third in developing countries for caloric contribution in human diets. Despite this importance, there has been a lack of genomic studies and resources for sweetpotato. We have constructed a BamHI BAC library in the pECBAC1 vector with an average insert size of 120 kb. This library is an essential resource for sweetpotato genomic studies and for the cloning of agriculturally and nutritionally important genes. A pair of degenerate PCR primers were designed based upon the on the P-loop and membrane motif sequences from five cloned resistance genes: RPS2, RPM1, N, L6, and Cre. Bands of the expected size have been observed at 500-bp and 700-bp from sweetpotato PCR amplifications. These bands have been cloned into pGEM-T easy vector and sequenced. The R gene candidate clones are being used as probes for BAC library screening and for the identification of coding and regulatory regions of resistance genes.