Legumes (Fabaceae or Leguminosae) are unique among cultivated plants for their ability to carry out endosymbiotic nitrogen fixation with rhizobial bacteria, a process that takes place in a specialized structure known as the nodule. Legumes belong to one of the two main groups of eurosids, the Fabidae, which includes most species capable of endosymbiotic nitrogen fixation 1. Legumes comprise several evolutionary lineages derived from a common ancestor 60 million years ago (Mya). Papilionoids are the largest clade, dating nearly to the origin of legumes and containing most cultivated species 2. Medicago truncatula (Mt) is a long-established model for the study of legume biology. Here we describe the draft sequence of the Mt euchromatin based on a recently completed BAC-assembly supplemented with Illumina-shotgun sequence, together capturing ~94% of all Mt genes. A whole-genome duplication (WGD) approximately 58 Mya played a major role in shaping the Mt genome and thereby contributed to the evolution of endosymbiotic nitrogen fixation. Subsequent to the WGD, the Mt genome experienced higher levels of rearrangement than two other sequenced legumes, Glycine max (Gm) and Lotus japonicus (Lj). Mt is a close relative of alfalfa (M. sativa), a widely cultivated crop with limited genomics tools and complex autotetraploid genetics. As such, the Mt genome sequence provides significant opportunities to expand alfalfa’s genomic toolbox.
We analyzed 6,749 lines tagged by the gene trap vector pGA2707. This resulted in the isolation of 3,793 genomic sequences flanking the T-DNA. Among the insertions, 1,846 T-DNAs were integrated into genic regions, and 1,864 were located in intergenic regions. Frequencies were also higher at the beginning and end of the coding regions and upstream near the ATG start codon. The overall GC content at the insertion sites was close to that measured from the entire rice (Oryza sativa) genome. Functional classification of these 1,846 tagged genes showed a distribution similar to that observed for all the genes in the rice chromosomes. This indicates that T-DNA insertion is not biased toward a particular class of genes. There were 764, 327, and 346 T-DNA insertions in chromosomes 1, 4 and 10, respectively. Insertions were not evenly distributed; frequencies were higher at the ends of the chromosomes and lower near the centromere. At certain sites, the frequency was higher than in the surrounding regions. This sequence database will be valuable in identifying knockout mutants for elucidating gene function in rice. This resource is available to the scientific community at http://www.postech.ac.kr/life/pfg/risd.Insertional mutagenesis is one of the most useful methods for analyzing gene function. When foreign DNA is inserted into a gene, it not only creates a mutation but also tags the affected gene, facilitating its isolation and characterization (Azpiroz-Leehan and Feldmann, 1997). Transposons and T-DNA have been used most widely as an insertional mutagen (Mathur et al., 1998;Wisman et al., 1998; Krysan et al., 1999;Parinov et al., 1999;Speulman et al., 1999;Tissier et al., 1999). It is believed that T-DNA insertion is a random event and that the inserted sequences are stable through multiple generations (Azpiroz- Leehan and Feldmann, 1997;Parinov and Sundaresan, 2000). Insertional mutant pools have been constructed in Arabidopsis and used for functional analysis of a number of genes (Feldmann, 1991; Koncz et al., 1992; Azpiroz-Leehan and Feldmann, 1997; Bechtold and Pelletier, 1998; Krysan et al., 1999; Galbiati et al., 2000;Parinov and Sundaresan, 2000; Bouché and Bouchez, 2001;Sessions et al., 2002;Szabados et al., 2002). The procedure for T-DNA insertional mutagenesis has also been applied to rice (Oryza sativa) using the Agrobacterium tumefaciensmediated transformation method (Hiei et al., 1994). Jeon et al. (2000) have reported the construction of over 20,000 T-DNA-tagged rice lines. A T-DNA insertional mutagen can be modified to trap a gene by inserting a reporter gene, such as gus (-glucuronidase), next to the T-DNA border (Sundaresan et al., 1995; Jeon et al., 2000;Springer, 2000). Approximately 5% to 10% of the mutagenized lines are GUS positive, demonstrating the efficiency of this gene-trapping system (Chin et al., 1999; Jeon et al., 2000).Completion of the genome sequencing for both Arabidopsis and rice has provided new reverse genetic means for assigning biological functions to sequenced genes (Kumar...
This study was conducted to investigate the promoter methylation status of the p16, DAPK, CDH1, and TIMP-3 genes in primary cervical cancer and its correlation with clinicopathologic characteristics. Promoter methylation was evaluated using a methylation-specific polymerase chain reaction in 78 cervical cancer tissue specimens and 24 control, normal cervical tissue specimens. Clinicopathologic parameters were obtained from medical records, and the relationship between the discrete variables and the methylation status was evaluated. The frequencies of promoter methylation of p16, DAPK, CDH1, and TIMP-3 in cervical cancer were 57%, 44.9%, 52.6%, and 9%, respectively. Primary cervical cancer had significantly higher methylation frequencies for the p16 and DAPK promoters than did the control, normal cervix (P < 0.0001). The promoter methylation of TIMP-3 was significantly higher in adenocarcinoma than in squamous cell carcinoma (41.7% vs 3%, respectively, P = 0.0175). High-stage cancers exhibited an increased promoter methylation frequency for p16 (P = 0.0061). The promoter methylation of the p16 gene is a frequent event in cervical carcinogenesis and may have potential clinical application as a marker for the progression and prognosis of cancer.
-SF 6 (sulphur hexafluoride) is the gas most commonly used in high voltage power equipments. Due to the fact that SF 6 is a greenhouse gas with a very high global warming potential, alternatives have been searched for over the last years. This paper presents the fundamental insulation characteristics of SF 6 /N 2 mixtures(50% SF 6 _50%N 2 , 20%SF 6 _80%N 2 ) in sphere-plane and rod-plane configurations. The electrode gap spacing and the relative gas pressure were varied within the range of 10~50mm and of 3~7bar respectively. To get the 50% breakdown value under lightning impulse of both polarity and AC, the up and down method was applied. In addition, both mixtures were tested in earthing switch and disconnecting switch sections of GIS model. The test result is applied to make design criteria for mixtures.
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