• International Journal of Industrial Entomology and Biomaterials
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• 제4권2호
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• pp.155-162
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• 2002

Cytochrome P45O (CYP) gene has been known to play one of the most important roles in metabolizing the exogenous materials. In insect, CYP is particularly known to detoxify toxic materials by adding oxygen molecule to the hydrophobic region of the materials. Thus, CYP-dependent metabolism is associated with the adaptation of insect to host plant chemicals. This in turn is known to be one of the driving forces for CYP diversification. In the present study, we cloned seven gene fragments of CYP 4 (CYP4) family from the midgut of the beet armyworm, Spodoptera exigua, through RT.PCT, Sequence analysis of the product showed the gene fragment to contain an open reading frame of ~150 amino acids, consisted of ~450 bp. The cloned gene fragments contained typical, conserved regions found in CYP4 family. Pairwise comparison of the deduced amino acid sequences among seven clones ranged in divergence from 0% to 52.86% and resulted in five distinct clones. The other two clones were identical or differ by one amino acid respectively to the corresponding clone, although each differed by ten nucleotides. Analysis of correlation between GenBank-registered, full length CYP4 and the cloned fragments resulted in statistically significant relationship ($r^{2}$ = 0.96085; p < 0.001), suggesting utility of the partial sequences as such full-length sequences. Phylogenetic analysis of the clones with GenBank-registered insect and mammal CYP4 family sequences by parsimony and several distance methods subdivided the clones into two groups: tones belonging to CYP4S and the others to CYP4M families.

• BMB Reports
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• 제41권10호
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• pp.716-721
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• 2008

To explore the effects of deregulated expression of the EBNA1 binding protein 2 (EBP2) on cell growth, we generated human HEK293 stable clones constitutively expressing an EBP2-EGFP fusion protein. We found both RNA and protein levels of cyclin E1, a dominant oncoprotein, were elevated in the EBP2- EGFP stable clones. These findings were confirmed by flow cytometry bivariate analysis of cyclin expression versus DNA content. Moreover, the increase in p21 expression and the specific phosphorylation at Ser1981 of ATM and Ser15 of p53 were also observed in these stable clones, and these observations may explain the failure to observe an increase in Cdk2 kinase activity. In addition, after one year of passage culture, the EBP2-EGFP stable clones tended to lose 4 to 5 chromosomes per cell when compared to that of control cells. All of these findings provide a possible link between deregulated expression of EBP2 and tumor development.

• Fisheries and Aquatic Sciences
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• 제10권3호
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• pp.119-126
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• 2007

The analysis of expressed sequence tags (ESTs) is an efficient approach for gene discovery, expression profiling, and the development of resources for functional genomics. To analyze the transcriptome of the abalone Haliotis discus hannai, we conducted EST analysis using seven cDNA libraries made from gill, gut, hepatopancreas, skin, muscle, testis, and ovary. Redundant ESTs were assembled into overlapping contiguous sequences using the assembly program ICAtools. We found that the total 1,393 ESTs formed 135 clusters and 951 singletons, indicating that the overall redundancy of the library was 22%. Of the 1,393 clones, BLAST identified 1,278 clones (91.7%) as known genes; 115 clones (8.3%) did not match any previously described gene. Based on the major functions of their encoded proteins, the identified clones were classified into 16 broad categories. Sequence analysis revealed the presence of micro satellite-containing genes that may be valuable for further gene mapping studies. This study contributes to the identification of numerous EST clones that can be applied to further clarifying the genetics and developmental biology of abalone.

• Molecules and Cells
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• 제42권12호
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• pp.869-883
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• 2019

Interleukin (IL)-15 is an essential immune-modulator with high potential for use in cancer treatment. Natural IL-15 has a low biological potency because of its short half-life and difficulties in mass-production. IL-15Rα, a member of the IL-15 receptor complex, is famous for its high affinity to IL-15 and its ability to lengthen the half-life of IL-15. We have double-transfected IL-15 and its truncated receptor IL-15Rα into CT26 colon cancer cells to target them for intracellular assembly. The secreted IL-15:IL-15Rα complexes were confirmed in ELISA and Co-IP experiments. IL-15:IL-15Rα secreting clones showed a higher anti-tumor effect than IL-15 secreting clones. Furthermore, we also evaluated the vaccine and therapeutic efficacy of the whole cancer-cell vaccine using mitomycin C (MMC)-treated IL-15:IL-15Rα secreting CT26 clones. Three sets of experiments were evaluated; (1) therapeutics, (2) vaccination, and (3) long-term protection. Wild-type CT26-bearing mice treated with a single dose of MMC-inactivated secreted IL-15:IL-15Rα clones prolonged survival compared to the control group. Survival of MMC-inactivated IL-15:IL-15Rα clone-vaccinated mice (without any further adjuvant) exceeded up to 100%. This protection effect even lasted for at least three months after the immunization. Secreted IL-15:IL-15Rα clones challenging trigger anti-tumor response via CD4⁺ T, CD8⁺ T, and natural killer (NK) cell-dependent cytotoxicity. Our result suggested that cell-based vaccine secreting IL-15:IL-15Rα, may offer the new tools for immunotherapy to treat cancer.

• 대한미생물학회지
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• 제35권2호
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• pp.149-157
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• 2000

Mycobacterium tuberculosis is capable of growing and survival within macrophage. The purpose of this study was to identify the genes regulated by infection of mycobacteria in human monocytic THP-1 cells. We used the differential display reverse transcriptase polymerase chain reaction (DD RT-PCR) and nothern blot analysis to confirm the differentially expressed genes from THP-1 cells infected with live Mycobacterium tuberculosis H37Rv, heat-killed Mycobacterium tuberculosis H37Rv and live Mycobacterium bovis BCG. Among many up or down-regulated clones, 27 clones were sequenced and compared with known genes on GenBank. Thirteen of over-expressed clones from THP-1 cells infected with live Mycobacterium tuberculosis H37Rv were identical to human prothymosin alpha, eight were novel clones and six clones showed homology with Human ferritin H chain, Esherichia coli bgl, Mouse RNA-dependent EIF-2 alpha kinase, E. coli htrL, Hyaluronan receptor and T cell receptor. Our result suggests that Mycobacterium tuberculosis might regulate prothymosin alpha gene transcription in monocytic THP-1 cell.

• Journal of Microbiology
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• 제34권1호
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• pp.25-29
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• 1996

Since sequencing of randomly selected cDNA clones has been known to be a powerful approach to obtain information on gene expression pattern in specific cells or tissues, we have analyzed a 3'-directed cDNA library of vegetative mycelia of A. nidulans by single-pass sequencing of hundreds of randomly selected clones. Sequencing of 292 cDNA clones yielded 209 gene signatures (GSs) probably representing highly or lesser expressed genes in the vegetative mycelia. Among the 209 GSs, 25 (79 cDNA clones) appeared more than once and 184 only once. One GS appeared at a highest frequency of 6 times, 2 GSs5 times, 4 GSs 4 times, a GSs 3 times and 16 GSs twice. About 6.6% GSs comprizing of 13 GSs showed alternative polyadenylation. Among 23 redundant GSs, three were common in both mycelia and sexual organs, and 22 were probably mycelia-specific. Out of 209 GSs, 36 were identified in GenBank showing of 70% or greater similaritis. Only six GSs were for A. nidulans genes, and 13 GSs were of DNA or genes encoding cytoplasmic or organellar proteins. This pattern is similar to those in the human HepG2 cell line and in human colonic mucosa, although very few genes for nuclear proteins and for protein synthesis were in A. nidulans.

• 한국작물학회:학술대회논문집
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• 한국작물학회 1999년도 춘계 학술대회지
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• pp.162-163
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• 1999

The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.

• 한국자원식물학회:학술대회논문집
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• 한국자원식물학회 2001년도 추계정기 학술발표회 초록집
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• pp.25-32
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• 2001

Potatoes have been recognized for a long time as one of the major food crops as well as horticultural crops. Potato production as a table food has been decreased in developed countries, while it has been steadily increased in the third world countries for it importance as food source. It is a new trend to look for the food, not only as a feeding crop but also healthy food. It is also time for the potato producers to look for the potato having high economic value as found in medicinal plants. There are great diversities in potato species, indicating that valuable compounds can be found in different amounts, depending on potato species. We screened the cultivars, breeding clones, and germplasms based on the vitamin C, Vitamin E, antioxidant compounds, diverse sugar types, important amino acids, and other valuable compounds. We could select the breeding clones KC003, 98Wl17, 99j717, and Vally 8 (A group) due to their high levels of antioxidant compounds, and it can be said that most of the red and purple colored potato clones belong to the A group. In the contents of essential amino acids, ‘Taebook Valley’,‘Summer Valley’ and other breeding clones were found to be high in amount. We also made crosses between breeding clones with high biological function and low agronomic traits and low biological function with high quality in agronomic characteristics . The patterns of genetic trends of these offsprings in comparison with their parents will be reported as well. And the potential of using potato as antibody production of anti-cancer will be discussed.

• 한국산림과학회지
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• 제36권1호
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• pp.1-4
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• 1977

본연구(本硏究)는 생장(生長)이 우수(優秀)하여 선발(選拔)한 ${\times}$P. albaglandulosa 15 clone의 엽(葉)에 대(對)하여 과산화동위효소변이(過酸化同位酵素變異)를 전기영동법(電氣泳動法)에 의(依)거 조사(調査)하여 다음과 같은 결과(結果)를 얻었다. 15 clone의 총(總) band수(數)는 6~11본(本)이다. 그중(中) 활성(活性) band수(數)는 4~7본(本)이고 흔적(痕迹) band는 1~4본(本)으로서 어느 clone에서나 활성(活性) band가 다수(多數)히 출현(出現)하였다. 그리고 cathode측(側)은 anode보다 band의 출현(出現)이 어느 clone에서도 단조(單調)로웠다. 한편(便) 효소형(酵素型)이 각(各) clone마다 특이(特異)하여 clone시별(識別)을 가능(可能)케 하였으며 g와 1 band는 ${\times}$P. albaglandulosa의 고정(固定) band이다. 또한 ${\times}$P. albaglandulosa가 $F_1$인 까닭에 효소형(酵素型)의 유전적(遺傳的) 변이(變異)가 인정(認定)되었다.

• IMMUNE NETWORK
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• 제11권2호
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• pp.100-106
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• 2011

Background: Disparities of Minor H antigens can induce graft rejection after MHC-matched transplantation. H60 has been characterized as a dominant antigen expressed on hematopoietic cells and considered to be an ideal model antigen for study on graft-versus-leukemia effect. Methods: Splenocytes from C57BL/6 mice immunized with H60 congenic splenocytes were used for establishment of H60-specific CTL clones. Then the clones were characterized for proliferation capacity and cytotoxicity after stimulation with H60. Clone #14, #15, and #23 were tested for the TCR binding avidity to H60-peptide/$H-2K^b$ and analyzed for TCR sequences. Results: H60-specific CTL clones showed different levels of proliferation capacity and cytotoxic activity to H60-stimulation. Clones #14, #15, and #23 showed high proliferation activity, high cytotoxicity, and low activities on both aspects, respectively, and have TCRs with different binding avidities to H60-peptide/$H-2K^b$ with $t_{1/2}$ values of 4.87, 6.92, and 13.03 minutes, respectively. The TCR usages were $V{\alpha}12D-3-01+J{\alpha}11-01$ and $V{\beta}12-1-01+D{\beta}1-01+J2-7-01$ for clone #14, $V{\alpha}13D-1-02+J{\alpha}34-02$ and $V{\beta}13-1-02+D{\beta}2-01+J{\beta}2-7-01$ for clone #15, and $V{\alpha}16D+J{\alpha}45-01$ and $V{\beta}12-1-01+D{\beta}1-01+J{\beta}2-5-01$ for clone #23. Conclusion: The results will be useful for modeling GVL and generation TCR transgenic mouse.