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KMID : 0368420060490020133
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Journal of Plant Biology
2006 Volume.49 No. 2 p.133 ~ p.140
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Identification of differentially expressed genes involved in spine formation on seeds ofDaucus carota L. (carrot), using annealing control primer (ACP) system
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Park Jong-Sug
Kim In-Sun
Cho Moon-Soo
Park Shin
Park Sang-Gyu
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Abstract
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Carrot seeds normally have surface spines. The availability of a spineless mutant would be agronomically beneficial, eliminating the current efforts to remove those spines pre-sowing. Furthermore, the identification of spine-specific genes would provide insights into spine development in wild-type carrot seed. This effort could be facilitated through the use of an annealing control primer (ACP) system. Here, we employed a new and accurate reverse transcriptionpolymerase chain reaction (RT-PCR) that involves ACPs for identifying genes of interest. With these techniques, 11 expressed sequence tags (ESTs) were obtained for cloning and sequencing the genes that are differentially expressed in wild-type spiny seeds, but not in the spineless mutant. In all, 7 cDNAs exhibited significant sequence similarity with known genes from other species. These included cell wall-associated hydrolase, tail fiber assembly protein, transcriptional regulatory protein, berberine bridge enzyme, S-adenosyl methionine synthase, transketolase, and phenylalanyl t-RNA synthetase beta chain. Four other cDNA sequences had no significant identities with known genes. As revealed by RT-PCR, these genes regulate spine formation during the developmental stage. Our results suggest that PCR-based differential display RT-PCR techniques are a very useful tool for identifying spine-specific genes from carrot seeds.
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KEYWORD
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annealing control primer (ACP), carrot, differentially expressed gene (DEC), RT-PCR, spineless, spiny
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