Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy
Since their initial discovery, transposons have been widely used as mutagens for forward and reverse genetic screens in a range of organisms. The problems of high copy number and sequence divergence among related transposons have often limited the efficiency at which tagged genes can be identified....
| Main Authors: | , , , , , , , , |
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| Format: | Journal Article |
| Language: | Inglés |
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Public Library of Science
2014
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10568/42279 |
| _version_ | 1855526721423409152 |
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| author | Howard, Thomas P. Hayward, Andrew P Tordillos, Anthony Fragoso, Christopher A. Moreno, Maria A Tohme, Joseph M. Kausch, Albert P Mottinger, John P Dellaporta, Stephen L. |
| author_browse | Dellaporta, Stephen L. Fragoso, Christopher A. Hayward, Andrew P Howard, Thomas P. Kausch, Albert P Moreno, Maria A Mottinger, John P Tohme, Joseph M. Tordillos, Anthony |
| author_facet | Howard, Thomas P. Hayward, Andrew P Tordillos, Anthony Fragoso, Christopher A. Moreno, Maria A Tohme, Joseph M. Kausch, Albert P Mottinger, John P Dellaporta, Stephen L. |
| author_sort | Howard, Thomas P. |
| collection | Repository of Agricultural Research Outputs (CGSpace) |
| description | Since their initial discovery, transposons have been widely used as mutagens for forward and reverse genetic screens in a range of organisms. The problems of high copy number and sequence divergence among related transposons have often limited the efficiency at which tagged genes can be identified. A method was developed to identity the locations of Mutator (Mu) transposons in the Zea mays genome using a simple enrichment method combined with genome resequencing to identify transposon junction fragments. The sequencing library was prepared from genomic DNA by digesting with a restriction enzyme that cuts within a perfectly conserved motif of the Mu terminal inverted repeats (TIR). Paired-end reads containing Mu TIR sequences were computationally identified and chromosomal sequences flanking the transposon were mapped to the maize reference genome. This method has been used to identify Mu insertions in a number of alleles and to isolate the previously unidentified lazy plant1 (la1) gene. The la1 gene is required for the negatively gravitropic response of shoots and mutant plants lack the ability to sense gravity. Using bioinformatic and fluorescence microscopy approaches, we show that the la1 gene encodes a cell membrane and nuclear localized protein. Our Mu-Taq method is readily adaptable to identify the genomic locations of any insertion of a known sequence in any organism using any sequencing platform. |
| format | Journal Article |
| id | CGSpace42279 |
| institution | CGIAR Consortium |
| language | Inglés |
| publishDate | 2014 |
| publishDateRange | 2014 |
| publishDateSort | 2014 |
| publisher | Public Library of Science |
| publisherStr | Public Library of Science |
| record_format | dspace |
| spelling | CGSpace422792025-03-13T09:44:20Z Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy Howard, Thomas P. Hayward, Andrew P Tordillos, Anthony Fragoso, Christopher A. Moreno, Maria A Tohme, Joseph M. Kausch, Albert P Mottinger, John P Dellaporta, Stephen L. zea mays genomics dna sequence maize transposons genómica secuencia de adn maíz transposones Since their initial discovery, transposons have been widely used as mutagens for forward and reverse genetic screens in a range of organisms. The problems of high copy number and sequence divergence among related transposons have often limited the efficiency at which tagged genes can be identified. A method was developed to identity the locations of Mutator (Mu) transposons in the Zea mays genome using a simple enrichment method combined with genome resequencing to identify transposon junction fragments. The sequencing library was prepared from genomic DNA by digesting with a restriction enzyme that cuts within a perfectly conserved motif of the Mu terminal inverted repeats (TIR). Paired-end reads containing Mu TIR sequences were computationally identified and chromosomal sequences flanking the transposon were mapped to the maize reference genome. This method has been used to identify Mu insertions in a number of alleles and to isolate the previously unidentified lazy plant1 (la1) gene. The la1 gene is required for the negatively gravitropic response of shoots and mutant plants lack the ability to sense gravity. Using bioinformatic and fluorescence microscopy approaches, we show that the la1 gene encodes a cell membrane and nuclear localized protein. Our Mu-Taq method is readily adaptable to identify the genomic locations of any insertion of a known sequence in any organism using any sequencing platform. 2014-01-31 2014-09-09T17:03:13Z 2014-09-09T17:03:13Z Journal Article https://hdl.handle.net/10568/42279 en Open Access Public Library of Science Howard III, Thomas P.; Hayward, Andrew P.; Tordillos, Anthony; Fragoso, Christopher; Moreno, Maria A.; Tohme, Joseph M.; Kausch, Albert P.; Mottinger, John P.; Dellaporta, Stephen L.. 2014. Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy. PLoS ONE 9(1): e87053. |
| spellingShingle | zea mays genomics dna sequence maize transposons genómica secuencia de adn maíz transposones Howard, Thomas P. Hayward, Andrew P Tordillos, Anthony Fragoso, Christopher A. Moreno, Maria A Tohme, Joseph M. Kausch, Albert P Mottinger, John P Dellaporta, Stephen L. Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy |
| title | Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy |
| title_full | Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy |
| title_fullStr | Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy |
| title_full_unstemmed | Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy |
| title_short | Identification of the maize gravitropism gene lazy plant1 by a transposon-tagging genome resequencing strategy |
| title_sort | identification of the maize gravitropism gene lazy plant1 by a transposon tagging genome resequencing strategy |
| topic | zea mays genomics dna sequence maize transposons genómica secuencia de adn maíz transposones |
| url | https://hdl.handle.net/10568/42279 |
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