Submitted to: Wheat Genetics International Symposium Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 10/15/2003
Publication Date: 11/1/2003
Citation: LUO, M., THOMAS, C.S., DEAL, K.R., YOU, F., ANDERSON, O.D., GU, Y.Q., LI, W., KURAPARTHY, V., GILL, B.S., MCGUIRE, P.E. 2003. CONSTRUCTION OF CONTIGS OF AEGILOPS TAUSCHII GENOMIC DNA FRAGMENTS CLONED IN BAC AND BIBAC VECTORS. WHEAT GENETICS INTERNATIONAL SYMPOSIUM PROCEEDINGS. 1:293-296. Interpretive Summary: Wheat is the most widely grown crop worldwide and feeds one third of the world's population. However, wheat has a large and complex genome as compared to model plants such as Arabidopsis and rice. Because of its genome complexity, cloning and manipulating genes controlling important traits for crop improvement remains a great challenge in wheat. A high-resolution physical map for the wheat genome will provide the basis for implementing broad-scale improvement of wheat. In this study, large wheat DNA fragments were first cloned into bacterial artificial chromosome (BAC) libraries. Based on overlapping sequences detected by fingerprinting technique used in this study, these fragments were then assembled with a computer program into much larger fragments, representing large portion of the whole genome or called physical maps for the wheat genome. Such physical maps will greatly facilitate gene cloning in wheat.
Technical Abstract: A high-throughput, fully automated, multi-color fluorescent fingerprinting technique for large-insert genomic DNA clones was developed. The technique was used to fingerprint 200,000 genomic DNA fragments of Aegilops tauschii line genetically closely related to the D genome of Chinese Spring wheat. The fragments were cloned in BAC and BiBAC vectors. The fingerprints were edited with a computer program (GenoProfiler) we developed and assembled into contigs with the FPC program. A total of 11,271 contigs was obtained at Sulston score of 1e-30 and tolerance 0.4 bp. Of these, 126 have more than 100 clones, 284 have 51 to 100 clones, 3,732 have 10 to 50 clones, and 7,129 have 2 to 9 clones. Many contigs have been anchored to wheat maps by integrating into them RFLP markers and ESTs. A database including fingerprints, BAC and BiBAC clones, contigs, and integrated markers has been developed and is publicly accessible on http://wheatdb.ucdavis.edu/PhysicalMapping/.