Plant & Animal Genomes XIV Conference Plant & Animal Genomes XIV ConferenceJanuary 14-18, 2006
Town & Country Convention Center
San Diego, CA
Charles H. Opperman1 , Steven Lommel1 , Mark Burke1 , Joanna Carlson1 , Carol George1 , Sandy Gove1 , Steve Graham1 , T.D. Houfek1 , Sam Kalat1 , P.C. Little1 , Amy Lumpkin1 , Lauren Redman1 , T. Ross1 , Reenah Schaffer1 , Elizabeth Scholl1 , P.J. Stephens1 , Eric Windham1 , S.H. Zekanis1 , Nathan Lakey2 , Joey Bidell2 , Arief Budiman2
Although cultivated tobacco, Nicotiana tabacum, is of great economic significance, relatively little information exists on its genome structure and organization. N. tabacum has a very large genome size compared with other cultivated solanaceous plants. At approximately 4.5 billion base pairs, it is 1.5 times the size of the human genome. As with the human genome, the vast majority of these base pairs occur as highly repetitive non-coding sequence. We are employing a combination of strategies to identify a large percentage of genes in N. tabacum. We are employing a methyl filtration library approach to identify gene-rich regions in N. tabacum in order to expedite the gene discovery. To date, we have sequenced 1,400,000 lanes of a methyl filtered library and observe a >10-fold increase in gene discovery in filtered vs. non-filtered libraries. In addition, we constructed a BAC library (9.7-fold genome coverage) and initiated BAC-end sequencing as part of our physical mapping program. We have fingerprinted 75,000 BAC clones using the four-dye protocol and have sequenced 38 BAC clones to ~5X coverage. We have also performed EST sequencing from various Nicotiana libraries and, to date, have sequenced 80,000 ESTs. In addition to sequencing methyl filtered clones and BACs, we are sequencing 6 newly constructed cDNA libraries to increase the EST acquisitions. Results from these experiments will be discussed, as well as overall gene discovery rates and genome organization.