• Title/Summary/Keyword: Gene-for-Gene

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Targeted Polymeric Gene Delivery for Anti-angiogenic Tumor Therapy

  • Kim, Won-Jong;Kim, Sung-Wan
    • Macromolecular Research
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    • v.15 no.2
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    • pp.100-108
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    • 2007
  • Gene therapy has become a promising strategy for the treatment of genetically based diseases, such as cancer, which are currently considered incurable. A major obstacle in the field of cancer gene therapy is the development of a safe and efficient delivery system for therapeutic gene transfer. Non-viral vectors have attracted great interest, as they are simple to prepare, stable, easy to modify and relatively safe compared to viral vectors. In this review, an insight into the strategies developed for polyethylenimine (PEI)-based non-viral vectors has been provide, including improvement of the polyplex properties by incorporating hydrophilic spacer, poly(ethylene glycol) (PEG). Moreover, this review will summarize the strategies for the tumor targeting. Specifically, a targeted polymeric gene delivery system, PEI-g-PEG-RGD, will be introduced as an efficient gene delivery vector for tumor therapy, including its functional analysis both in vitro and in vivo.

Expression of Replication-Independent Chicken H3.3 Histone Gene without Introns

  • Son, Seung-Yeol;Hong, Bum-Shik
    • BMB Reports
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    • v.30 no.3
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    • pp.200-204
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    • 1997
  • We eliminated introns from replication independent chicken H3.3 histone gene using a H3.3 cDNA clone and a genomic H3.3 clone. After introduction into Rat 3 cells, we observed its pattern of expression by analyzing mRNA from different phases of the cell cycle. Even without introns, the H3.3 gene was expressed constitutively at a low level throughout the cell cycle. This indicates that the introns in the H3.3 gene are not responsible for the cell cycle-independent expression of the gene. This result contradicts previous reports that suggested their importance in cell cycle regulated expression. We believe that other regions of the gene, promoter, coding region, and/or 3'-end of the gene, are involved in its expression pattern.

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Sequence-specific interaction between ABD-B homeodomain and castor gene in Drosophila

  • Kim, Keon-Hee;Yoo, Siuk
    • BMB Reports
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    • v.47 no.2
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    • pp.92-97
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    • 2014
  • We have examined the effect of bithorax complex genes on the expression of castor gene. During the embryonic stages 12-15, both Ultrabithorax and abdominal-A regulated the castor gene expression negatively, whereas Abdominal-B showed a positive correlation with the castor gene expression according to real-time PCR. To investigate whether ABD-B protein directly interacts with the castor gene, electrophoretic mobility shift assays were performed using the recombinant ABD-B homeodomain and oligonucleotides, which are located within the region 10 kb upstream of the castor gene. The results show that ABD-B protein directly binds to the castor gene specifically. ABD-B binds more strongly to oligonucleotides containing two 5'-TTAT-3' canonical core motifs than the probe containing the 5'-TTAC-3' motif. In addition, the sequences flanking the core motif are also involved in the protein-DNA interaction. The results demonstrate the importance of HD for direct binding to target sequences to regulate the expression level of the target genes.

Analysis of Molecular Epidemiological Properties of Staphylococcus aureus Isolates from Domestic Animals and Human Patients by PCR (Polymerase Chain Reaction을 활용한 국내 동물과 사람환자에서 분리한 Staphylococcus aureus 분리주의 분자역학적 특성분석)

  • Woo Yong-Ku;Kim Shin
    • Korean Journal of Microbiology
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    • v.41 no.1
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    • pp.24-37
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    • 2005
  • This study was conducted to analyze the molecular epidemiological properties and to select the most efficient and reliable PCR method on 116 of Staphylococcus aureus (S. aureus) isolates from Korean cattle, black goat, pig, dog, chicken, mouse and also human clinical cases from hospital. The distribution patterns of SSG [species specific genes; coagulase (coa), protein A (spa), nuclease (nuc) and aroA (RsaI) gene] were analyzed by PCR method. Among the SSGs, the nuc-gene was found in all strains $(100\%)$ tested and followed by coa-gene $(87.9\%)$, spa-gene $(91.4\%)$ and aroA-gene $(26.7\%)$, in order. The genetic subtyping by RFLP method was performed on the coa [AluI] and aroA-gene [RsaI] PCR products. The mecA-gene PCR and PCR-RFLP techniques were chosen to detect and verify of MRSA strains. Only the human strains $(12.1\%)$ were detected the positive mecA-gene products (533 bp), which were divided into two specific bands [201 & 332 bp] by HhaI enzyme digestion. On coa-gene and spa-gene typing, coa-gene was typed with ten kinds of genotype and coa-3 type were determined as the most predominant genotype, while spa-gene was divided into eleven kinds of genotype and also spa-7 type were selected the most prevalent genotype based on their genetic variations. On the aroA and coa-gene subtyping by PCR-RFLP, aroA-gene products were discriminated with only seven types of genotype, while coa-gene products were further divided into an eleven genotype, respectively. In comparison of SID values of five PCR based typing methods, the coa-PCR-RFLP (SID0.894) was evaluated the most efficient and reliable tools and followed by coa-PCR (SID0.883) and aroA-PCR-RFLP (SID0.462), in order. In conclusion, we could determined that the coa-PCR-RFLP method was the most suitable genetic analysis tool for S. aureus and MRSA strains from domestic animals and humans.

PCR-mediated Recombination of the Amplification Products of the Hibiscus tiliaceus Cytosolic Glyceraldehyde-3-phosphate Dehydrogenase Gene

  • Wu, Linghui;Tang, Tian;Zhou, Renchao;Shi, Suhua
    • BMB Reports
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    • v.40 no.2
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    • pp.172-179
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    • 2007
  • PCR-mediated recombination describes the process of in vitro chimera formation from related template sequences present in a single PCR amplification. The high levels of genetic redundancy in eukaryotic genomes should make recombination artifacts occur readily. However, few evolutionary biologists adequately consider this phenomenon when studying gene lineages. The cytosolic glyceraldehyde-3-phosphate dehydrogenase gene (GapC), which encodes a NADP-dependent nonphosphorylating glyceraldehyde-3-phosphate dehydrogenase in the cytosol, is a classical lowcopy nuclear gene marker and is commonly used in molecular evolutionary studies. Here, we report on the occurrence of PCR-mediated recombination in the GapC gene family of Hibiscus tiliaceus. The study suggests that recombinant areas appear to be correlated with DNA template secondary structures. Our observations highlight that recombination artifacts should be considered when studying specific and allelic phylogenies. The authors suggest that nested PCR be used to suppress PCRmediated recombination.

Catabolite Repression of the Bacillus stearothermophilus $\beta$-Xylosidase Gene (xylA) in Bacillus subtilis

  • Cho, Ssang-Goo;Choi, Yong-Jin
    • Journal of Microbiology and Biotechnology
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    • v.8 no.1
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    • pp.21-27
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    • 1998
  • The xylA gene of Bacillus stearothermophilus encoding the major ${\beta}$-xylosidase was previously cloned and sequenced. In the present study we examined the regulation of the cloned xylA gene expression in Bauillus subtilis MW15 carrying the xylA::aprA fusion plasmids. The induction of the fused xylA gene expression remained uninfluenced by any of the carbon sources tested but the gene expression was repressed about 2-3 fold in the presence of glucose. Two CRE-like sequences (CRE-1: nucleotides + 124 to +136 and CRE-2: +247 to +259) were recognized within the reading frame region of the xylA gene. The deletion experiments showed that the CRE-2 sequence had a role in catabolite repression (CR) as a true CRE of the xylA gene, but the CRE-1 had no effect on CR of the xylA gene expression. Surprisingly, the deletion of the CRE- 1 sequence reduced about 2~3 fold of the expression of the xylA fused gene. The repression ratios of the xylA gene expression were estimated to be about 0.4 from the assay of subtilisin activity, and about 0.3 at the level of transcription by determining the amounts of xylA transcripts in B. subtilis. While, the level of CR of the xylA gene was assessed to be about l0-fold in previous work when the relative amounts of the xylA transcripts were measured in B. stearothermophilus.

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Hybrid Fungal Genome Annotation Pipeline Combining ab initio, Evidence-, and Homology-based gene model evaluation

  • Min, Byoungnam;Choi, In-Geol
    • 한국균학회소식:학술대회논문집
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    • 2018.05a
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    • pp.22-22
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    • 2018
  • Fungal genome sequencing and assembly have been trivial in these days. Genome analysis relies on high quality of gene prediction and annotation. Automatic fungal genome annotation pipeline is essential for handling genomic sequence data accumulated exponentially. However, building an automatic annotation procedure for fungal genomes is not an easy task. FunGAP (Fungal Genome Annotation Pipeline) is developed for precise and accurate prediction of gene models from any fungal genome assembly. To make high-quality gene models, this pipeline employs multiple gene prediction programs encompassing ab initio, evidence-, and homology-based evaluation. FunGAP aims to evaluate all predicted genes by filtering gene models. To make a successful filtering guide for removal of false-positive genes, we used a scoring function that seeks for a consensus by estimating each gene model based on homology to the known proteins or domains. FunGAP is freely available for non-commercial users at the GitHub site (https://github.com/CompSynBioLab-KoreaUniv/FunGAP).

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Function and use of silkworm dpp gene (누에 유래의 dpp 유전자의 기능 및 유전체 연구를 통한 식의약용 신소재로서 활용)

  • Park, Seung Wong
    • Journal of Sericultural and Entomological Science
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    • v.54 no.1_2
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    • pp.1-5
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    • 2016
  • The dpp gene originated from the silkworms is an important gene that is well conserved in the genome of humans, cattle, rodents, poultry and Drosophila. The dpp gene belonging to the TGF-beta (Transforming Growth Factor-beta) superfamily is known to play an important role in several developmental stages. The $TGF-{\beta}$ gene family is a genetically well-conserved and playing an important role gene family in various species such as determining cell proliferation and differentiation, apoptosis and cell fate. In this review, we have confirmed the following studies data. The recent studies on the silkworm dpp gene have confirmed for the first time the biological functions such as promoting osteogenesis activity. In addition, previous data shows that dpp have developmental functions such as morphogenetic materials at the blastophyllum stage, induction of the mesoblast at the late embryonic stage and involved in the proliferation and morphogenesis of imaginal disc in adult development. We found the splice variant of the dpp gene originated from the wildtype silkworm by using comparative genomics. It has provided important data for basic research based on genetics studies of these processes may promote a better understanding of evolution. Silkworm is a medicinal insect and is approved for its safety. It is used as a natural antibiotic for promoting growth as a medical material, a health functional food, and a feed additive. Therefore, it is necessary to present various data to obtain more value of functional insect.

Correlation between Expression Level of Gene and Codon Usage

  • Hwang, Da-Jung;Han, Joon-Hee;Raghava, G P S
    • Proceedings of the Korean Society for Bioinformatics Conference
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    • 2004.11a
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    • pp.138-149
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    • 2004
  • In this study, we analyzed the gene expression data of Saccharomyces cerevisiae obtained from Holstege et al. 1998 to understand the relationship between expression level and nucleotide sequence of a gene. First, the correlation between gene expression and percent composition of each type of nucleotide was computed. It was observed that nucleotide 'G' and 'C' show positive correlation (r ${\geq}$ 0.15), 'A' shows negative correlation (r ${\approx}$ -0.21) and 'T' shows no correlation (r ${\approx}$ 0.00) with gene expression. It was also found that 'G+C' rich genes express more in comparison to 'A+T' rich genes. We observed the inverse correlation between composition of a nucleotide at genome level and level of gene expression. Then we computed the correlation between dinucleotides (e.g. AA, AT, GC) composition and gene expression and observed a wide variation in correlation (from r = -0.45 for AT to r = 0.35 for GT). The dinucleotides which contain 'T' have wide range of correlation with gene expression. For example, GT and CT have high positive correlation and AT have high negative correlation. We also computed the correlation between trinucleotides (or codon) composition and gene expression and again observed wide range of correlation (from r = -0.45 for ATA r = 0.45 for GGT). However, the major codons of a large number of amino acids show positive correlation with expression level, but there are a few amino acids whose major codons show negative correlation with expression level. These observations clearly indic ate the relationship between nucleotides composition and expression level. We also demonstrate that codon composition can be used to predict the expression of gene in a given condition. Software has been developed for calculating correlation between expression of gene and codon usage.

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