Plant & Animal Genome IX Conference Plant & Animal Genome IX Conference
Poster: Large Insert Libraries, Gene Isolation, Etc.
P02_07.html
A physical map integrated with the developed genetic maps will provide a readily used platform for effectively large-scale gene cloning, EST mapping, target marker development and genome sequencing. BAC fingerprinting and contig assembly has provided powerful tools to rapidly generate physical maps with bacteria-based large-insert clones. Using the fingerprinting technologies, we previously developed a genome-wide BAC-based physical map of rice and a genome-wide BAC/BIBAC-based physical map of Arabidopsis. To develop a BAC/BIBAC-based physical map of the entire soybean genome, we previously constructed two large-insert soybean (Glycine max) cv. Forest DNA libraries in the plant transformation-competent binary vector pCLD04541 with Hind III and Bam HI, respectively. Recently, we have developed a Forest BAC library in the Eco RI site of the pECBAC1 vector. This new BAC library contains 38,400 clones and has an average insert size of 157 kb. Because these three libraries were constructed with three different enzymes in two different vector systems, they are complementary to each other and thus, have a truly genome coverage. A total of about 80,000 BACs and BIBACs have been randomly selected from the libraries and fingerprinted by the DNA sequencing electrophoresis-based restriction fingerprinting method. The fingerprints of over 20,000 clones have been put into computer workstations, edited and created into FPC database. The progress in the construction of the physical map will be reported. The BAC/BIBAC-based physical map is being integrated with the existing genetic maps and the large-scale functional genomics research of soybean. The results will promise to revolutionize the current structural, functional and comparative genomics of this important crop.