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- Title
Construction and Identification of Bacterial Artificial Chromosome Library for 0-613-2R in Upland Cotton.
- Authors
Jian-Mei Yin; Wang-Zhen Guo; Tian-Zhen Zhang
- Abstract
A bacterial artificial chromosome (BAC) library containing a large genomic DNA insert is an important tool for genome physical mapping, map-based cloning, and genome sequencing. To isolate genes via a map-based cloning strategy and to perform physical mapping of the cotton genome, a high-quality BAC library containing large cotton DNA inserts is needed. We have developed a BAC library of the restoring line 0-613-2R for isolating the fertility restorer (Rf1) gene and genomic research in cotton (Gossypium hirsutum L.). The BAC library contains 97 825 clones stored in 255 pieces of a 384-well microtiter plate. Random samples of BACs digested with the NotI enzyme indicated that the average insert size is approximately 130 kb, with a range of 80–275 kb, and 95.7% of the BAC clones in the library have an average insert size larger than 100 kb. Based on a cotton genome size of 2 250 Mb, library coverage is 5.7 × haploid genome equivalents. Four clones were selected randomly from the library to determine the stability of the BAC clones. There were no different fingerprints for 0 and 100 generations of each clone digested with NotI and HindIII enzymes. Thus, the stability of a single BAC clone can be sustained at least for 100 generations. Eight simple sequence repeat (SSR) markers flanking the Rf1 gene were chosen to screen the BAC library by pool using PCR method and 25 positive clones were identified with 3.1 positive clones per SSR marker. (Managing editor: Li-Hui Zhao)
- Subjects
ARTIFICIAL chromosomes; BACTERIAL artificial chromosomes; DNA; COTTON; PLANT genetic engineering; PLANT gene mapping; PLANT genetics -- Technique
- Publication
Journal of Integrative Plant Biology, 2006, Vol 48, Issue 2, p219
- ISSN
1672-9072
- Publication type
Article
- DOI
10.1111/j.1744-7909.2006.00169.x-i1