• Title/Summary/Keyword: gene library

Search Result 663, Processing Time 0.02 seconds

TNF-induced genes and Proteins

  • 이태호
    • Proceedings of the Korean Society of Applied Pharmacology
    • /
    • 1993.11a
    • /
    • pp.17-20
    • /
    • 1993
  • As a step toward a more complete understanding of the molecular actions of TNF, we prepared a cDNA library from TNF-treated human FS-4 fibroblasts and used differential hybridization to identify cDNA clones corresponding to mRNAs enriched in TNF-treated eells. In Quiescent FS-4 cells n induces an increase in the level of some mRNAs within 20 to 30 min. Some of these immediate-early response mRNAs are elevated only transiently for about 30 to 120 min, e. g., c-fos and c-myc (Lin and Vilcek,1987) or the transcription factor IRF-1 (Fujita et al.1989). Such immediate-early gene products may be important for the activation of other genes, but their transient induction suggests that they are not the actual effector molecules responsible for the phenotypic changes induced by TNF. We chose a 3-h incubation with W because we were seeking cDNAs corresponding to messages that are more stably elevated after TNF treatment. Indeed, the results shown in Figure 8 and 9 indicate that all of the mRNAs corresponding to the eight TSG cDNAs isolated remained significantly elevated after 16h of continuous treatment with TNF, and their kinetics of induction were clearly different from those of the immediate-early response mRNAs such as c-fos, c-myc or IRF-1. Nevertheless, only the induction of TSG-21 (collagenase) and TSG-27 (stromelysin) nNAs was completely inhibited by cycloheximide and the induction of TSG-37 (metallothionein-II) was reduced in the presence of this inhibitor of protein synthesis. Induction of the other five TSG mRNAs by TNF was completelyresistant to cycloheximide, suggest ins that no protein intermediate is needed for the upregulation of these mRNAs.

  • PDF

Platform of Hot Pepper Defense Genomics: Isolation of Pathogen Responsive Genes in Hot Pepper (Capsicum annuum L.) Non-Host Resistance Against Soybean Pustule Pathogen (Xanthomonas axonopodis pv. glycines)

  • Lee, Sang-Hyeob;Park, Do-Il
    • The Plant Pathology Journal
    • /
    • v.20 no.1
    • /
    • pp.46-51
    • /
    • 2004
  • Host resistance is usually parasite-specific and is restricted to a particular pathogen races, and commonly is expressed against specific pathogen genotypes. In contrast, resistance shown by an entire plant species to a species of pathogen is known as non-host resistance. Therefore, non-host resistance is the more common and broad form of disease resistance exhibited by plants. As a first step to understand the mechanism of non-host plant defense, expressed sequence tags (EST) were generated from a hot pepper leaf cDNA library constructed from combined leaves collected at different time points after inoculation with non-host soybean pustule pathogen (Xanthomonas axonopodis pv. Glycines; Xag). To increase gene diversity, ESTs were also generated from cDNA libraries constructed from anthers and flower buds. Among a total of 10,061 ESTs, 8,525 were of sufficient quality to analyze further. Clustering analysis revealed that 55 % of all ESTs (4685) occurred only once. BLASTX analysis revealed that 74% of the ESTs had significant sequence similarity to known proteins present in the NCBI nr database. In addition, 1,265 ESTs were tentatively identified as being full-length cDNAs. Functional classification of the ESTs derived from pathogen-infected pepper leaves revealed that about 25% were disease- or defense-related genes. Furthermore, 323 (7%) ESTs were tentatively identified as being unique to hot pepper. This study represents the first analysis of sequence data from the hot pepper plant species. Although we focused on genes related to the plant defense response, our data will be useful for future comparative studies.

Detection of Ref-1 (Redox factor-1) Interacting Protein Using the Yeast Two-hybrid System (Yeast two-hybrid system을 이용한 Ref-1 (redox factor-1) 결합 단백질의 분리 및 동정)

  • 이수복;김규원;배문경;배명호;정주원;안미영;김영진
    • Journal of Life Science
    • /
    • v.14 no.1
    • /
    • pp.26-31
    • /
    • 2004
  • Redox factor-1 (Ref-1), known as a redox regulator, controls the DNA binding of AP-1 and is activated in HT29 colon cancer cells by hypoxia in vitro. REF-1 also increases tile DNA binding affinity of Hypoxia-inducible Factor-lalpha$ (HIF-lalpha$), HIF-like Factor (HLF) and early growth response-1 (Egr-1) which induce expression of the genes involved in angiogenesis, so that we speculate that REF-1 may play a role in hypoxia-induced angiogenesis. In this research we tried to detect novel proteins interacting with REF-1 using Yeast two-hybrid system using full-length REF-1 cDNA as bait. As result of such screening we detected 3 positive clones. DNA sequencing and GeneBank search revealed that one of the clones contained the same sequences as M.musculus cDNA for tioredoxin.

Molecular Cloning and Identification of Novel Genes, Gomsin, Characteristically Expressed in Snailfish, Liparis tanakae (꼼치에서 특징적으로 발현되는 새로운 유전자 곰신의 분리 및 동정)

  • 송인선;이석근;손진기
    • Development and Reproduction
    • /
    • v.6 no.1
    • /
    • pp.7-16
    • /
    • 2002
  • In order to obtain the specific genes of snailfish a subtracted cDNA library was constructed, and analysed by sequencing and GenBank search. Among them C90-171 clone was turned out to be genes showing low homology and nonredundant genes. This novel clone was named Gomsin(C90-171). Gomsin was shown to be intensely expressed in the epithelial cells, some mesenchymal cells, and sheaths of muscle bundles in the result of immunohistochemistry. In the cross reaction assay of Gomsin antibody against various human tissues, the Gomsin was strongly expressed in the ductal and acinar cells of salivary glands, which was similar to the expression patterns of proline-rich proteins(PRPs) of human. The antibody raised against the Gomsin was clearly cross-reacted with human salivary PRPs and also recombinant proteins of human PRPs in the Western blot and immunoprecipitation analysis. Contrast to the salivary PRPs, the Gomsin was not easily degraded in the mixed saliva, but rapidly attacked on the cultured keratocytes in vitro. The simulated protein structure of Gomsin was similar to the whorled pattern of PRPs, even though the amino acid sequence of Gomsin was quite different from those of PRPs. These data suggest that the Gomsin is a characteristic matrix protein in the skin and body of snailfish, which is also utilized for the tissue protection in the similar way to the PRPs of human muco-secretory organs.

  • PDF

ABCB1 Polymorphisms and Imatinib Response in Chronic Myeloid Leukemia Patients: A Systematic Review and Meta-analysis (ABCB1 유전적 다형성이 만성 골수성 백혈병 환자의 Imatinib 치료 반응에 미치는 영향: 체계적 문헌고찰 및 메타분석)

  • Ha, Hye Min;Chun, Pusoon
    • YAKHAK HOEJI
    • /
    • v.60 no.3
    • /
    • pp.118-127
    • /
    • 2016
  • A growing number of studies have demonstrated that ABCB1 gene polymorphisms are associated with the variability of responses to imatinib. However, the effects of ABCB1 polymorphisms on imatinib response in chronic myeloid leukemia (CML) are inconsistent. The aim of the present study was to clarify the associations between ABCB1 polymorphisms and imatinib response in CML. A systematic literature review was performed. The databases of PubMed, Embase, and Cochrane Library were searched for all published studies from inception to December 2015. The following terms were used with functions of 'AND' and 'OR': 'chronic myeloid leukemia', 'CML', 'ABCB1', 'MDR1', 'polymorphism', 'SNP', and 'imatinib'. Using the Review Manager 5, odds ratios (ORs) were pooled to estimate the effect of ABCB1 polymorphisms on imatinib response in CML. The pooled analysis showed that ABCB1 2677 G allele was significantly associated with poor response to imatinib in African and Asian patients (GG vs TT, OR: 0.32, p<0.0001; GG+GT vs TT, OR: 0.44, p=0.0005). In subgroup analyses, African patients carrying ABCB1 1236 C allele exhibited higher risk for worse response, whereas Asian patients with 1236 C allele showed better response (CC+CT vs TT, OR: 0.41, p=0.008 for African; OR: 1.65, p=0.03 for Asian). There was no association between C3435T polymorphisms and imatinib response in African, Asian, and Caucasian CML patients.

Development of SSR markers for genetic mapping of Korean ginseng and authentication of Korean ginseng cultivars

  • Kim, Nam-Hoon;Choi, Hong-Il;Jung, Ju-Yeon;Choi, Beom-Soon;Ahn, In-Ok;Lee, Joon-Soo;Yang, Tae-Jin
    • Proceedings of the Plant Resources Society of Korea Conference
    • /
    • 2010.10a
    • /
    • pp.11-11
    • /
    • 2010
  • The Korean ginseng, Panax ginseng C. A. Meyer is a popular medicinal herb in Araliaceae. Genetic map in crops provides valuable information for breeding, genetic and genomic researches. However, little information is available for construction of genetic map in ginseng. Up to now, we have produced large amounts of expressed sequence tags (ESTs) from four ginseng cultivars (37Mb, 49Mb, 39Mb, 47Mb from Gopoong, Gumpoong, Chunpoong and Yunpoong respectively using pyrosequencing technique and 5Mb from normalized full-length cDNA library of Chunpoong) to obtain comprehensive information of gene expression, and constructed EST database including ESTs from public database. Till now, we designed 261 SSR primer sets using EST sequences and identified 106 intergenic polymorphic markers. And 44 of the 106 showed polymorphisms among panax ginseng cultivars. Among 44 markers, 27 SSR polymorphic markers were inspected to 51 $F_2$ population from Yunpoong x Chunpoong, which showed good at the fitness of Mendellian segregation ratio 1:2:1. To enrich the number of markers, and thus construct high resolution genetic map which can be used as frame map for further genome sequencing. we are planning to develop large scale EST-derived SNP markers which are available in the F2 population. This study provides genetic information as well as foundation for ginseng researches such as genetics, genomics, breeding, and the final goal for whole genome sequencing. This study was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea (Grant No. 609001-051SB210).

  • PDF

IVET-based Identification of Virulence Factors in Vibrio vulnificus MO6-24/O

  • Lee, Ko-Eun;Bang, Ji-Sun;Baek, Chang-Ho;Park, Dae-Kyun;Hwang, Won;Choi, Sang-Ho;Kim, Kum-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • v.17 no.2
    • /
    • pp.234-243
    • /
    • 2007
  • Vibrio vulnificus is an opportunistic pathogen that causes septicemia in humans. To identify the genes associated with its pathogenicity, in vivo expression technology (IVET) was used to select genes specifically expressed in a host, yet not significantly in vitro. Random lacZ-fusions in the genome of V vulnificus strain MO6-24/O were constructed using an IVET vector, pSG3, which is a suicide vector containing promoterless-aph and -lacZ as reporter genes. A total of ${\sim}18,000$ resulting library clones were then intraperitoneally injected into BALB/c mice using a colony forming unit (CFU) of $1.6{\times}10^6$. Two hours after infection, kanamycin was administered at $200{mu}g$ per gram of mouse weight. After two selection cycles, 11 genes were eventually isolated, which were expressed only in the host. Among these genes, VV20781 and VV21007 exhibiting a homology to a hemagglutinin gene and tolC, respectively, were selected based on having the highest frequency. When compared to wild-type cells, mutants with lesions in these genes showed no difference in the rate of growth rate, yet a significant decrease in cytotoxicity and the capability to form a biofilm.

Enhanced Production of Bacterial Cellulose in Komagataeibacter xylinus Via Tuning of Biosynthesis Genes with Synthetic RBS

  • Hur, Dong Hoon;Choi, Woo Sung;Kim, Tae Yong;Lee, Sang Yup;Park, Jin Hwan;Jeong, Ki Jun
    • Journal of Microbiology and Biotechnology
    • /
    • v.30 no.9
    • /
    • pp.1430-1435
    • /
    • 2020
  • Bacterial cellulose (BC) has outstanding physical and chemical properties, including high crystallinity, moisture retention, and tensile strength. Currently, the major producer of BC is Komagataeibacter xylinus. However, due to limited tools of expression, this host is difficult to engineer metabolically to improve BC productivity. In this study, a regulated expression system for K. xylinus with synthetic ribosome binding site (RBS) was developed and used to engineer a BC biosynthesis pathway. A synthetic RBS library was constructed using green fluorescent protein (GFP) as a reporter, and three synthetic RBSs (R4, R15, and R6) with different strengths were successfully isolated by fluorescence-activated cell sorting (FACS). Using synthetic RBS, we optimized the expression of three homologous genes responsible for BC production, pgm, galU, and ndp, and thereby greatly increased it under both static and shaking culture conditions. The final titer of BC under static and shaking conditions was 5.28 and 3.67 g/l, respectively. Our findings demonstrate that reinforced metabolic flux towards BC through quantitative gene expression represents a practical strategy for the improvement of BC productivity.

The Acceptance of Electronic Journals among Undergraduate Students in Korea (대학생의 전자저널 수용에 관한 연구)

  • Chang, Hye-Rhan;Jung, Eu-Gene
    • Journal of the Korean Society for information Management
    • /
    • v.28 no.1
    • /
    • pp.237-262
    • /
    • 2011
  • The purpose of this study is to understand the acceptance of the electronic journals among undergraduate students. Using a survey questionnaire, this study collected the data collected from 813 students taking 11 courses offered in five different universities. The levels of electronic jounal and its service awareness appeared to be low as 63.7% and 59.9% respectively. Only 43.8% of the respondents report their experience of using Korean e-journals. There is a statistically significant difference in use among college groups and students' major of study. E-journals are mostly used off-campus and within the library, using simple search techniques. In spite of difficulties in search and gathering information, the satisfaction level marked above average. Reasons of nonuse are lack of opportunity and knowledge. However, 93.8% of nonusers report a willingness to use the e-journals in the future. LIS students show higher e-journal awareness and use experience compared to non-LIS students. Yet, the levels of using simple search techniques and satisfaction show little difference from those of the non-LIS students. The findings of the study suggest implications to promote the acceptance of e-journals among college students.

Molecular Cloning of a cDNA Encoding a Cathepsin D Homologue from the Mulberry Longicorn Beetle, Apriona germari

  • Kim, Seong-Ryul;Yoon, Hyung-Joo;Park, Nam-Sook;Lee, Sang-Mong;Moon, Jae-Yu;Jin, Byung-Rae;Sohn, Hung-Dae
    • International Journal of Industrial Entomology and Biomaterials
    • /
    • v.3 no.2
    • /
    • pp.121-126
    • /
    • 2001
  • A cDNA encoding a cathepsin D homologue was cloned from a cDNA library of the mulberry longicorn beetle, Apriona germari. Sequence analysis of the cDNA encoding the cathepsin D homologue of A. germari revealed that the 1,158 bp cDNA has an open reading frame of 386 amino acid residues. The deduced protein sequence of the A. germari cathepsin D homologue shows high homology with cathepsin D in insects, Aedes aegypti (68.2% amino acid similarity) and Drosophila melanogaster (67.2% amino acid similarity). Two aspartic residues and six cystein residues in the A. germari cathepsin D homologue are present at identical locations in all of the other catepsins D. Unlike cathepsins D in two insect species, A. gemari cathepsin D homologue appears to have two putative glycosylation sites, rather than one. Phylogenetic analysis revealed the A. germari cathepsin D homologue is more closely related to insect cathepsins D than to the other animal cathepsins D. Northern blot analysis suggests that A. germari cathepsin D homologue gene is expressed in most if not all, body tissues.

  • PDF