Sunday, May 3, 2009
4-08
Random Shear BAC Cloning: An Optimized Genomic Tool for Improving Biomass and Bioenergy Feedstocks
David Mead, Rosa Ye, Svetlana Jasinovica, Ronald Godiska, and Cheng-Cang Wu. Lucigen, 2120 W Greenview Dr, Middleton, WI 53562
Bacterial artificial chromosome (BAC ) libraries, BAC -based physical mapping, and genome sequencing are critical for understanding the molecular basis of phenotypic variation. This knowledge can be used to accelerate breeding and engineering of new plant varieties, exploiting natural genetic diversity to increase yield and improve resistance to biotic and abiotic stresses. BAC libraries built with conventional vectors and methods are inherently biased. Numerous gaps exist in all of the physical and sequencing maps of eukaryotic genomes, for example Arabidopsis and rice. We have developed techniques to construct unbiased, randomly-sheared BAC libraries with large inserts (>100 kb) as well as a unique transcription-free BAC vector. We have used random shear BAC libraries to close numerous gaps in conventional deep-coverage BAC libraries for many International Genome Projects, including Arabidopsis, rice, potato, tomato, soybean, barley, etc. Efficient whole genome sequencing of biomass/bioenergy plants from a single Random Shear BAC library has been also demonstrated recently (e.g., for oil palm and Jatropha curcas). Lucigen’s Random Shear BAC service is publicly available (see www.lucigen.com) and provides a novel genomics tool to efficiently characterize and understand plant cell wall synthesis and deconstruction.