An integrated system for large-scale functional analysis of the genome sequence in Arabidopsis thaliana /
A genome sequence map of the plant model species, Arabidopsis thaliana ecotype Columbia, has been recently developed, except for the centromere regions. To facilitate the gap closure of the sequence map and large-scale functional analysis of the genome sequence experimentally, we have developed a g...
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| Format: | Thesis Book |
| Language: | English |
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[Place of publication not identified] :
[publisher not identified] ;
2001.
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| Online Access: | http://proxy.library.tamu.edu/login?url=http://proquest.umi.com/pqdweb?did=728908041&sid=1&Fmt=2&clientId=2945&RQT=309&VName=PQD |
| Summary: | A genome sequence map of the plant model species, Arabidopsis thaliana ecotype Columbia, has been recently developed, except for the centromere regions. To facilitate the gap closure of the sequence map and large-scale functional analysis of the genome sequence experimentally, we have developed a genome-wide, BAC/BIBAC-based, integrated physical, genetic and sequence map of A. thaliana ecotype Columbia. The map consists of 194 contigs, covering 126 Mb. Of the 194 nuclear DNA BAC/BIBAC contigs, 120 were anchored to the five chromosomes of A. thaliana, spanning about 114 Mb. The accuracy of this map was verified by using the information from the public physical and sequence maps and library screening with numerous mapped RFLP markers. The integrated map has been successfully used in the closure of gaps in the sequence map. Furthermore, since the BIBACs were designed to transfer plants by the Agrobacterium-mediated transformation method, this map has provided a readily used platform for large-scale map-based cloning of Arabidopsis genes and functional analysis of the genome sequence. This study also demonstrated that the DNA sequencing gel-based restriction fingerprinting method is reliable and powerful for genome-wide physical mapping with large-insert BACs and /or BIBACs. To facilitate positional cloning of Arabidopsis genes and functional analysis of its genome sequence, we have also developed a large-insert, plant-transformation-competent BIBAC library of A. thaliana ecotype Landsberg. This library was constructed in the Bam HI site of the binary vector BIBAC2 and contains 11,520 clones with an average insert size of 162 kb. Clone analysis showed that 2,074 clones of the library had no inserts, 289 were originated from the chloroplast genome and 24 from the mitochondrial genome. Therefore, the library contains 9,132 clones covering 11.4x haploid genomes of A. thaliana. Further characterization showed that seven clones, on average, could be identified with each DNA marker, 0.8% of the 9,132 clones contained 180-bp repeats, 0.67% contained 5S rDNA, and 0.82% contained 18S-25S rDNA. This library represents the first large-insert genomic DNA and plant-transformation-competent BIBAC library of the Landsberg genome and provides valuable resources for positional cloning, functional analysis, and comparative genomics research in A. thaliana. |
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| Item Description: | Vita. "Major Subject: Plant Physiology". |
| Physical Description: | ix, 85 leaves : illustrations ; 28 cm. Issued also on microfiche from University Microfilm Inc. |
| Bibliography: | Includes bibliographical references (leaves 73-84). |