• Journal of Pharmaceutical Investigation
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• v.34 no.5
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• pp.351-362
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• 2004

The basic concept underlying gene therapy is that human diseases may be treated by the transfer of genetics material into specific cells of a patient in order to correct or supplement defective genes responsible for disease development. There are several systems that can be used to transfer foreign genetic material into the human body. Both viral and non-viral vectors are developed and evaluated for delivering therapeutic genes. Viral vectors are biological systems derived from naturally evolved viruses capable of transferring their genetics materials into host cells. However, the limitations associated with viral vectors, in terms of their safety, particularly immunogenecity, and their limited capacity of transgenic materials, have encouraged researchers to increasingly focus on non-viral vectors as an alternative to viral vectors. Although non-viral vectors are less efficient than viral ones, they have the advantages of safety, simplicity of preparation and high gene encapsulation capability. This article reviews the most recent studies highlighting the advantages and the limitation of gene delivery systems focused on non-viral systems compared to viral systems.

• Environmental Analysis Health and Toxicology
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• v.24 no.2
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• pp.107-117
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• 2009

Metal pollution of aquatic ecosystems is a problem of economic and health importance. Information regarding molecular responses to metal exposure is sorely needed in order to identify potential biomarkers. To determine the effects of heavy metals on chironomids, the full-length cDNA of alcohol dehydrogenase (ADH3) from Chironomus riparius was determined through molecular cloning and rapid amplification of cDNA ends (RACE). The expression of ADH3 was analyzed under various cadmium and copper concentrations. A comparative and phylogenetic study among different orders of insects and vertebrates was carried out through analysis of sequence databases. The complete cDNA sequence of the ADH3 gene was 1134 bp in length. The sequence of C. riparius ADH3 shows a low degree of amino acid identity (around 70%) with homologous sequences in other insects. After exposure of C. riparius to various concentrations of copper, ADH3 gene expression significantly decreased within 1 hour. The ADH3 gene expression was also suppressed in C. riparius after cadmium exposure for 24 hour. However, the effect of cadmium on ADH3 gene expression was transient in C. riparius. The results show that the suppression of ADH3 gene by copper exposure could be used as a possible biomarker in aquatic environmental monitoring and imply differential toxicity to copper and cadmium in C. riparius larvae.

• Journal of Plant Biotechnology
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• v.36 no.1
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• pp.45-52
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• 2009

In order to understand the molecular interactions that occur between rice and the rice blast fungus during infection, we previously identified a number of rice blast fungal elicitor-responsive genes from rice (Oryza sativa cv. Milyang 117). Here, we report the cloning and characterization of the rice fungal elicitor-inducible gene Oshin1 (GenBank Accession Number AF039532). Sequence analysis revealed that the Oshin1 cDNA is 1067 bp long and contains an open reading frame encoding 205 amino acid residues. The Oshin1 gene shows considerable sequence similarity to the tobacco hin1 and hin2 genes. The predicted Oshin1 protein has a cysteine-rich domain at the N-terminus and is rich in leucine, serine, and alanine residues. Southern blot analysis suggests that Oshin1 gene is a member of a small gene family in the rice genome. To examine the expression of Oshin1, Northern blot analysis was conducted. Expression of the Oshin1 transcript is rapidly induced in suspension-cultured rice cells treated with fungal elicitor, salicylic acid or hydrogen peroxide. In addition, Oshin1 transcript levels are rapidly increased by treatment with $Ca^{2+}$/A23187. The expression of Oshin1 was also elevated in 3-week old leaf tissues upon ethephon application or fungal elicitor treatment. Our results suggest that the Oshin1 gene is involved in plant defense responses to environmental stresses.

• Journal of Plant Biology
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• v.35 no.2
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• pp.165-171
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• 1992

In order to study regulation of rbcL gene expression, rbcL gene of chloroplast DNA (Cp DNA) from maize was cloned. Cp DNA was isolated from intact chloroplast and digested with BamHI. BamHI 9 fragment of Cp DNA containing rbcL gene was ligated to pUC19 and transformed into E. coli DH5a. This recombinant plasmid was named pRLYSl. pRLYSl was hybridized with a part of rbcL gene from rice and digested with restriction enzyme BamHI, HindIIl, and PstI. From these results, it was confirmed that pRLYS1 contains intact rbcL gene and orientation of BamHI 9 fragment of Cp DNA in pRLYS1 was determined.rmined.

• Journal of Mushroom
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• v.9 no.2
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• pp.53-58
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• 2011

In order to confirm the presence of putative glucoamylase gene in Tricholoma matsutake genome, the genomic DNA was prepared from T. matsutake NBRC30773 strain and was used as template to clone the glucoamylases gene (TmGlu1). We obtained the nucleotide sequence of TmGlu1 and its franking region. The coding region (from ATG to stop codon) is 2,186 bp. The locations of exons and introns were determined from the nucleotide sequences of 3'- and 5'-RACE PCR and RT-PCR products. On the other hand, to investigate the relationship between composition of medium and glucoamylase expression, we checked the expression level of glucoamylase gene by realtime reverse transcription PCR and measurement of glucoamylase enzyme activity. It was found that enzyme activity of glucoamylase was very low in different medium. Expression of glucoamylases gene appeared to not be affected by different carbon source.

• Macromolecular Research
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• v.17 no.1
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• pp.19-25
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• 2009

In order to enhance the gene delivery efficiency and decrease the cytotoxicity of polyplexes, we synthesized Solutol-g-PEI by conjugating polyethyleneimine (PEI) to Solutol (polyoxyethylene (10) stearate), and evaluated its efficiency as a possible nonviral gene carrier candidate. Structural analysis of synthesized polymer was performed by using $^1H$-NMR. Gel retardation assay, particle sizes and zeta potential measurement confirmed that the new gene carrier formed a compact complex with plasmid DNA. The complexes were smaller than 150 nm, which implicated its potential for intracellular delivery. It showed lower cytotoxicity in three different cell lines (Hela, MCF-7, and HepG2) than PEI 25 kDa. pGL3-lus was used as a reporter gene, and the transfection efficiency was in vitro measured in Hela cells. Solutol-g-PEI showed much higher transfection efficiency than unmodified PEI 25 kDa.

• Plant Resources
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• v.1 no.1
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• pp.6-12
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• 1998

The Fp1 gene, originally isolated from red pepper seedlings, encode the iron storage protein, and have a high homology with ferritin genes at DNA and amino acid level. In order to determine ferritin protein expression in vegetative tissue. Fp1 gene was constructed in plant expression vector(PIG12IHm) and introduced in red pepper(var. Bukang, Chungyang and Kalag-Kimjang 2) via Agrobacterium tumefaciensmediated transformation. After selection on MS media containing Kanamycin(Km), putatively selected transformants were confirmed by amplification of selectable marker gene(Fp1 and NPII) by polymerase chain reaction. Northern blot showed that transcripts of Fp1 gene were detected in mature leaves of the plants. In A6, A7 and A8 and A14 of transgenic plants, transcript of Fp1 gene was increased seven-fold to eight-fold than other transgenic plants. Also the proteins obtained from leaves of transgenic plants were immunologically detected by Western blot using rabbit anti-ferritin polyclonal antibody. The expression protein appeared as strong band of apparent mass of 23.5kDa. suggesting the iron accumulation in transgenic red pepper plants.

• Proceedings of the Botanical Society of Korea Conference
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• 1996.07a
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• pp.10-18
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• 1996

Recently, we have reported a rice MADS-box gene, OsMADS1, as a molecular factor triggering flower formation; this has been well studied in a heterologous system (Chung et al., 1994). In order to study whether the OsMADS1 homolog exists in other plant species, the OsMADS1 cDNA was used as a probe to screen a tobacco cDNA library, and a potential homolog, NtMADS3, was isolated. Sequence analysis revealed that the gene shares 56.1% identity in whole amino acids with OsMADS1. Like OsMADS1, the NtMADS3 gene starts to express at a very early stage of flower development, and the expression continues up to flower maturation. In the tobacco flower, the gene is expressed in whorl 2,3 and 4, corresponding to the petal, stamen, and carpel, respectively. Upon ectopic expression in the homologous system, NtMADS3 caused a trasition from inflorescence shoot meristem into floral meristem, reducing flowering time dramatically. These phenotypes strongly suggest the NtMADS3 gene is the OsMADS1 homolog of tobacco. Hybrids between the OsMADS1 and the NtMADS3 plants were also generated. The hybrids flowered even earlier than these two transgenic plants. The detailed studies are discussed here.

• Journal of Microbiology
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• v.44 no.2
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• pp.217-225
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• 2006

Kaposi's sarcoma-associated herpesvirus (KSHV) RTA transcription factor is recruited to its responsive elements through interaction with RBP-Jk that is a downstream transcription factor of the Notch signaling pathway that is important in development and cell fate determination. This suggests that KSHV RTA mimics cellular Notch signal transduction to activate viral lytic gene expression. Here, I demonstrated that unlike other B lymphoma cells, KSHV -infected primary effusion lymphoma BCBL1 cells displayed the constitutive activation of ligand-mediated Notch signal transduction, evidenced by the Jagged ligand expression and the complete proteolytic process of Notch receptor I. In order to investigate the effect of Notch signal transduction on KSHV gene expression, human Notch intracellular (hNIC) domain that constitutively activates RBP-Jk transcription factor activity was expressed in BCBL1 cells, TRExBCBL1-hNIC, in a tetracycline inducible manner. Gene expression profiling showed that like RTA, hNIC robustly induced expression of a number of viral genes including KS immune modulatory gene resulting in downregulation of MHC I and CD54 surface expression. Finally, the genetic analysis of KSHV genome demonstrated that the hNIC-mediated expression of KS during viral latency consequently conferred the downregulation of MHC I and CD54 surface expression. These results indicate that cellular. Notch signal transduction provides a novel expression profiling of KSHV immune deregulatory gene that consequently confers the escape of host immune surveillance during viral latency.

• Korean Journal of Plant Resources
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• v.20 no.6
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• pp.524-529
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• 2007

We carried out to study the function of ArgE in transgenic rice plants, which were confirmed by PCR analysis and hygromycin selection. Transgenic rice plants were with selectable marker gene(HPT) inserted in genome of the rice. Southern analysis with hpt probe confirmed by two restriction enzymes that copy numbers of the selectable gene was introduced into the plant genome. We displayed that the relationship between drought stress and ArgE gene with the overexpressing rice plants. From this result, we observed that the degree of leaves damage has no difference in control and transgenic lines. The total RNAs were extracted from 6 weeks-seedling in normal condition in order to examine their expression levels with ArgE-overexpressed transgenic rice. In particular, expression patterns of genes encoding enzymes involved in abiotic stress, including drought and salt stresses. OsGF14a and OsSalt were investigated by reverse transcription-PCR(RT-PCR). Expression levels of the OsSalt gene decreased significantly in transgenic rice plants compared to control plant. However, ion leakage measurement did not demonstrate any leaves damage change between control and ArgE transgenic plants exposure to mannitol treatment. These results suggest that expression of the ArgE is not involved in tolerance for drought stress in rice but may playa role of signaling networks for salt-induced genes.