Construction and characterization of a bacterial artificial chromosome library of the Indica rice [Oryza sativa subsp. Indica] Kasalath
1999
Miyazaki, C. (Japan Science and Technology Corp., Ohmura, Nagasaki) | Saito, A.
We constructed a bacterial artificial chromosome (BAC) library for the indica rice variety Kasalath using a new BAC vector, pCHR8, suited for plasmid rescue, containing nine unique restriction sites in a multicloning site. This Kasalath BAC library consists of about 65,000 clones with an average insert size of 106 kb using HindIII partially digested DNA fragments. It represents approximately sixteen rice haploid genome equivalents which are sufficient for the full coverage of the rice genome with BAC clone contigs. The library was constructed from only a single ligation mixture with high transformation efficiency. The quality of this BAC library was ascertained by successfully retrieving the BAC clones for seven rice RFLP markers. We could isolate eleven individual BAC clones using RFLP markers that linked to the lethal factor locus for segregation distortion on rice chromosome 3. The content of the chloroplast and mitochondrial DNA in this library was estimated to be 2.8% and 0.4%, respectively. These results demonstrate that this Kasalath BAC library should be suitable for map-based cloning and physical mapping of the rice genome
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