Identification of chloroplast DNA insertions in nuclear chromosomes of maize B73 line using the FISH procedure

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Identification of chloroplast DNA insertions in nuclear chromosomes of maize B73 line using the FISH procedure

Please use this identifier to cite or link to this item: http://hdl.handle.net/10355/2084

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dc.contributor.author Donnelly, Laura en
dc.contributor.author Westgate, Leah en
dc.contributor.author Meyer, Louis J., 1978- en
dc.contributor.author Alberts, Patrice en
dc.contributor.author Newton, Kathleen J. en
dc.contributor.author Birchler, James A. (James Arthur), 1950- en
dc.date 2005 en
dc.date.accessioned 2009-07-14T21:56:56Z en
dc.date.available 2009-07-14T21:56:56Z en
dc.date.issued 2005 en
dc.identifier.uri http://hdl.handle.net/10355/2084 en
dc.description Abstract only available en
dc.description.abstract It is known that chloroplast DNA can incorporate itself into the nuclear genome of plants. However, the sites of chloroplast (ct) DNA integration into chromosomes of maize have not yet been analyzed. This project is the first attempt to find the location of the ctDNA on the maize chromosomes. Fluorescent in situ hybridization is a technique that has proved useful in karyotyping and chromosomal mapping in maize. The FISH procedure is being used in this study to discover the location of the ctDNA in the nuclear genome of the inbred line B37. In order to develop ctDNA “probes” for FISH analysis, we have used the polymerase chain reaction (PCR) to produce fragments of ctDNA. Primers were chosen to amplify fragments of 10 kb or larger. The amplified DNAs were purified and labeled with fluorescent dyes and these probes were subsequently hybridized to chromosomes. The probes recognize and bind to the corresponding DNA sequences within the chromosomes. Root tip cells were used to prepare the slides for hybridization. Because the cells are collected during the metaphase stage of division, the chromosomes are compact and more easily visible. Chromosomes that contain ctDNA can be detected using a compound microscope with fluorescent attachments. The location of the ctDNA on the chromosomes is made visible by the fluorescent labeling of the probe. Eight of eleven regions of the chloroplast genome of the B73 line have been specifically amplified and have been observed under the microscope for FISH analysis. This information will contribute to an understanding of the extent and mechanism of transfer of organellar genomes to the nucleus. en
dc.description.sponsorship MU Monsanto Undergraduate Research Fellowship en
dc.language en_US en
dc.language.iso en_US en
dc.publisher University of Missouri--Columbia. Office of Undergraduate Research en
dc.relation.ispartof 2005 Summer Undergraduate Research and Creative Achievements Forum (MU) en
dc.source.uri http://undergradresearch.missouri.edu/forums-conferences/abstracts/abstract-detail.php?abstractid= en
dc.subject chloroplast DNA en
dc.subject nuclear chromosomes of maize en
dc.subject fluorescent in situ hybridization technique (FISH) en
dc.subject organellar genomes en
dc.title Identification of chloroplast DNA insertions in nuclear chromosomes of maize B73 line using the FISH procedure en
dc.type Presentation en
dc.contributor.meetingname Summer Undergraduate Research and Creative Achievements Forum (2005 : University of Missouri--Columbia) en
dc.contributor.corporatename University of Missouri-Columbia. Office of Undergraduate Research en
dc.relation.ispartofcommunity University of Missouri-Columbia. Office of Undergraduate Research. Undergraduate Research and Creative Achievements Forum


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