VitE (tocotrienols and tocopherols) are micronutrients with antioxidant properties synthesized by photosynthetic bacteria and plants that play important roles in animal and human nutrition. A new mutant line, T1001-1, was isolated from in vitro mutagenized population by ionizing radiation and shown to have increased VitE contents. The total VitE content was 26% increased in the T1001-1 mutant seeds compare with cv. Dongan (wild-type). In addition, we showed that the mutant confers retarded seedling growth during the early seedling growth stage in rice. To study the molecular mechanism of VitE biosynthesis, we used the rice microarray to identify genes that are upor down-regulated in T1001-1 mutant. In addition, we identified differentially regulated pathway using MapMan analysis, which provides deep insight into changes in transcript and metabolites. Our results enhanced the transcription of genes involved in starch and lipid metabolism in T1001-1 mutant. To identify the molecular mechanisms of the events involving transcription factors in tocopherol accumulation, we compared the expression patterns of transcription factors. The AP2-EREBP, WRKY, C2H2 transcription factor were up-regulated, whereas the MYB family was down-regulated in T1001-1 mutant. Our results demonstrate change of important transcript in high level of VitE accumulating rice mutant.

Manganese () is a trace element that is essential for normal physiology, and is predominantly obtained from food. Several lines of evidence, however, demonstrated that overexposure to exerts serious neurotoxicity, immunotoxicity and developmental toxicity, particularly in male. The present study aimed to evaluate the effect of 0, 1.0, 3.3, and 10 mg/kg/day doses of on the reproductive organs in the immature female rats. Rats (PND 22; S.D. strain) were exposed to () dissolved in drinking water for 2 weeks. The animals were sacrificed on PND 35, then the tissues were immediately removed and weighed. Histological studies were performed using the uteri tissue samples. Serum LH and FSH levels were measured with the specific ELISA kits. Body weights of the experimental group animals were not significantly different from those of control group animals. However, ovarian tissue weights in 1 mg and 3.3 mg dose groups were significantly lower than those of control animals (p<0.05 and p<0.01, respectively). Uterine tissue weights of 3.3 mg dose groups were significantly lower than those of control animals (p<0.01), while the 1 mg dose and 10 mg dose failed to induce any change in uterine weight. Similarly, only 3.3 mg dose could induce the significant decrease in the oviduct weight compared to the control group (p<0.05). Non-reproductive tissues such as adrenal and kidney failed to respond to all doses of exposure. The uterine histology revealed that the exposure could affect the myometrial cell proliferation particularly in 3.3 mg dose and 10mg dose group. Serum FSH levels were significantly decreased in 1mg dose and 10 mg groups (p<0.05 and p<0.01, respectively). In contrast, treatment with 1 mg dose induced a significant increment of serum LH level (p<0.05). The present study demonstrated that exposure is capable of inducing abnormal development of reproductive tissues, at least to some extent, and altered gonadotropin secretions in immature female rats. Combined with the well-defined actions of this metal on GnRH and prolactin secretion, one can suggest the might be a potential environmental mediator which is involved in the female pubertal process.

Ionizing radiation is known to cause chromosomal alterations such as inversions and deletions and affects gene expression within the plant genome. To monitor the genome-wide transcriptome changes by ionizing radiation, we used rice Affimetrix GeneChip microarray to identify genes that are up- or down regulated by gamma-ray (200 Gy, 60Co source), cosmic-ray and ion beam (40 Gy, 220 MeV carbon ion). The overall expression patterns between gamma-ray and ion beam were similar but cosmic-ray was regulated differently. Combined results from all 3 radiations identified 27 up-regulated genes and 188 down regulated genes. These results mean the induction of similar mechanism changes in treatments of gamma ray and ion beam. However the different expression in treatment of cosmic-ray might be due to the other environmental conditions. Among the commonly up- or down- regulated genes, we chose highly up- or down- regulated several genes and confirmed its regulation in response to ionizing radiation exposure by RT-PCR analysis. Moreover, we showed that specific co-expression networks of candidate radio marker genes by ARACNE algorithm. Our results present profiles of gene expression related to different ionizing radiation and marker gene to predict sensitivity to ionizing radiation, such as GS (glutelin subunit) and FBX322.

Discovery, identification, and informatics of low molecular weight peptide are extensively rising in the field of proteomics research. In this study, we analyzed protein profiles to discover peptide based biomarker for twelve different soybean seeds with three different agronomic types using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS). For optimization of SELDI-TOF MS in soybean seed proteome analysis, four different extraction buffers were tested with urea solubilization buffer, thiourea/urea solubilization buffer, phenol extraction buffer, and modified trichloroacetic acid (TCA)/acetone precipitation/urea solubilization extraction buffer. Two different type of ProteinChip arrays, cation exchange (CM10) and anion exchange (Q10), applied to profile peptides. Among the four different extraction buffers, phenol extraction was selected to protein extraction methodology. Numbers of detected peak cluster in twelve soybean seeds were 125 at CM10 and 90 at Q10 array in the mass range from 2 to 40 kDa. Among them, 82 peak clusters at CM10 and 33 peak clusters at Q10 array showed significantly different peak clusters at p<0.00004 (CM10) and p<0.00005 (Q10) among twelve different soybean cultivars. Moreover, 29 peak clusters at CM10 and 17 peak clusters at Q10 array were detected in all cultivars as an ‘universally existed peptide’. In comparison with three different agronomic types, total of 55 peak clusters (CM10) and 23 peak clusters (Q10) were significantly different peak clusters at p<0.00004 and p<0.0001, respectively. In these probability levels, soybean seeds were well discriminated into different cultivar and different type with each other. Also we could find several specific peptide biomarkers for agronomic type.

The canine major histocompatibility complex (MHC) is referred to dog leukocyte antigens (DLA), which is known to be the most polymorphic genetic system in canine species. Many cloned dogs have been produced since Snuppy, first cloned dog, there was no research about genetic identity of MHC among cloned animals. Recently in Lee’s group, two non-transgenic cloned beagles (BG1, 2) were produced by somatic cell nuclear transfer (SCNT) using fetal fibroblast (BF). Also, four transgenic cloned beagles (Ruppy 1-3, 5) were generated using transgenic BF transfected with Red fluorescent protein (RFP) gene. We hypothesize that non-transgenic (BG1, 2) and transgenic (Ruppy 1-3, 5) cloned beagles derived from identical donor cells have the same immunological genetic characteristic except for RFP gene insertion in the genome. Thus, the aim of this study is to confirm the immunological identity of DLA class II in cloned beagles produced using same nuclear donor cell. Genomic DNA was extracted from blood of BG1, BG2, Ruppy 1, 2, 3 and 5. Genomic DNA of normal two control beagle, no correlation with BF was also investigated for rulling out the possibility that beagles were inbred. Forward and reverse primers used for DLA-DQA1 and DQB1 respectively were DQAF: 5’-TAAGGTTCTTTTCTCCCTCT-3’ and DQAR: 5’-GGACAGATTCAGTGAAGAGA-3’ DQBR:5’-CTCACTGGCCCGGCTGTCTC-3’ and DQBR: 5’-CACCTCGC CGCTGCAACGTG-3’. Polymerase Chain Reaction (PCR) products were purified, sequenced directly using the Big Dye Terminator kit. Sequencing analysis was performed on an automated 3730xl DNA analyzer. In experiment 1, sequence of DLA-DQ alpha 1 (DQA1) and DLA-DQ beta 1 (DQB1) exon 2, hypervariabel region, was compared in BG1 and BG2. Experiment 2 also compared the sequence of DQA1 and DQB1 among Ruppy 1, 2, 3 and 5. Experimental 3 compared sequence of DQA1 and DQB1 among all cloned dogs (BG1, BG2 and Ruppy 1-3, 5). As a result, BG1 and BG2 have same allele for DQA1 and DQB1 as we expected. They share DQA1*00101 and DQB1*02901 in experiment 1. In experiment 2, Ruppy 1, 2, 3 and 5 also have identical DQA1*00101 and DQB1*02901 allele. No discrimination between transgenic dogs and cloned dogs was seen in DQA1 and DQB1 Allele in experiment 3. DQA1, DQB1 allele was identified as *00101 and *02901 in all dogs. We provided the allele identity of DQA1and DQB1 in cloned beagles, which can be used as preliminary data for immunological related studies. In conclusion, transgenic cloned dogs despite of red fluorescent protein genes being inserted in their nuclear DNA were immunologically compatible with non-transgenic cloned dogs. We demonstrated that cloned beagles produced using identical nuclear donor were immunologically compatible.

A resveratrol synthase (RS) gene was isolated from peanut (Arachis hypogaea, L. cv. Jinpoong) plants. This gene was placed under the control of the cauliflower mosaic virus 35S promoter (CaMV35S) and introduced into two Korean varieties of potato (Solanum tuberosum L. cvs. Jasim and Jowon) plants by Agrobacterium-mediated gene transfer. Putative transformants were screened by PCR with primers designed from CaMV 35S promoter, NOS terminator and RS gene. Most of selected transgenic potato plants showed the amplification of expected fragments by PCR of genomic DNA with gene-specific primers, while they were absent in untransformed control plants. Expression of the resveratrol synthase gene was also examined by northern blot analysis. The transformants showed a band which was lacking in the control plant, confirming that the introduced gene is transcribed into mRNA in the transformants. The strength of the band, which reflected the level of mRNA expression, differed among the individual transformants. Among the transformants obtained, the highest trans-resveratrol content in the transgenic young leaves of purple-fleshed "Jashim" was 2.11 μgg-1 fresh weight and that in the microtubers in vitro of purple fleshed "Jashim" was 8.31 μgg-1 fresh weight. This amount of resveratrol may have a positive biological effect on human health.

We performed field trials to assess levels of changes in intrinsic properties and resistance against soft rot of the potato cultivar ‘Irish Cobbler’ upon the introduction of the Shiva-1 gene. Each five lines, transformed with Shiva-1 gene controlled by the PAL5 promoter (P) and by the CaMV 35S promoter (E) were evaluated in the field. In based on evaluation of 10 defined morphological characteristics, all the transgenic clones of both lines proved to be true to type. When five agronomic characteristics were taken into account as well, all the transgenic lines except E8 were considered to be true to type. According to the result of northern blot analysis, seven (P1, P3, P4, P6, E10, E12, and E16) transgenic clones could be distinguished clearly from corresponding untransformed clones. But in three lines (P8, E5, and E8), no hybridization signal was detected. There seems to be positive correlation between the levels of resistance to soft rot and the transgenic expression at mRNA levels in P lines. But in the case of E lines, however, there doesn’t seem to be any correlation between the levels of disease resistance and mRNA expression