• 대한임상검사과학회지
• /
• 제43권3호
• /
• pp.89-97
• /
• 2011

The Human Papilloma Virus (HPV) infection has been strongly associated with pathogenesis of uterine cervix carcinoma. HPV type 16, a causative agent of uterine cervix carcinoma, encodes the E6 and E7 oncogenes, expression of which is pivotal for malignant transformation and maintenance of malignant phenotypes. To develop a gene therapy for HPV-related carcinoma, We investigated the effect of E6 short-interfering RNA (E6 siRNA) on the expression of this oncogene and on the growth of HPV 16-related uterine cervix carcinoma cells. SiHa cells, a uterine cervix carcinoma cell line, which contain a single copy of HPV 16 integrated in the chromosome and express the E6 and E7 oncogenes. Before 24 hr of transfection, cells were seeded and transfected with control plasmid or E6 siRNA-expressing plasmid. The mRNA was analysed by reverse transcriptase polymerase chain reaction (RT-PCR). The cell growth rate was investigated by MTT method. The E6 mRNA level in SiHa cells was decreased in HPV 16 E6 siRNA-expression vector transfected cells and a decrease in the growth of these cells was also observed. From these results. it is evident that E6 siRNA played a role in suppression of growth of SiHa cells and has a fair chance as a candidate for gene specific therapy for HPV related uterine cervix carcinoma.

• 한국응용약물학회:학술대회논문집
• /
• 한국응용약물학회 2006년도 Spring Conference
• /
• pp.77-90
• /
• 2006

Beta-amyloid peptide (A$\beta$) is a major component of senile plaques and its aggregation is considered to play a critical role in pathogenesis of Alzheimer's disease (AD). Aggregation of A$\beta$ could result from both increased synthesis and decreased degradation of A$\beta$. Our laboratory is interested in understanding the mechanism of A$\beta$ degradation in brain. Recently our laboratory identified a bacterial gene (SKAP) from Streptomyces sp KK565 whose protein product has an activity to cleave A$\beta$ and thus reduce the A$\beta$-induced neurotoxicity. The sequence analysis showed that this gene was closely related to aminopeptidase. Maldi-Tof analysis showed that the recombinant SKAP protein expressed in E. coli cleaves both A$\beta$ 40 and A$\beta$ 42 at the N-terminal of A$\beta$ while an aminopeptidase from Streptomyces griseus (SGAP) cleaves at the C-terminal. We also identified a mammalian homolog of SKAP and the recombinant mammalian protein expressed in Sf-9 insect cells showed a similar proteolytic activity to SGAP, cutting A$\beta$ at the C-terminus. I well discuss the detailed mechanism of the enzyme action and its functional implication in AD.

• 한국균학회소식:학술대회논문집
• /
• 한국균학회 2014년도 추계학술대회 및 정기총회
• /
• pp.9-9
• /
• 2014

Null mutants generated by targeted gene replacement are frequently used to reveal function of the genes in fungi. However, targeted gene deletions may be difficult to obtain or it may not be applicable, such as in the case of redundant or lethal genes. Constitutive expression system could be an alternative to avoid these difficulties and to provide new platform in fungal functional genomics research. Here we developed a novel platform for functional analysis genes in Magnaporthe oryzae by constitutive expression under a strong promoter. Employing a binary vector (pGOF1), carrying $EF1{\beta}$ promoter, we generated a total of 4,432 transformants by Agrobacterium tumefaciens-mediated transformation. We have analyzed a subset of 54 transformants that have the vector inserted in the promoter region of individual genes, at distances ranging from 44 to 1,479 bp. These transformants showed increased transcript levels of the genes that are found immediately adjacent to the vector, compared to those of wild type. Ten transformants showed higher levels of expression relative to the wild type not only in mycelial stage but also during infection-related development. Two transformants that T-DNA was inserted in the promotor regions of putative lethal genes, MoRPT4 and MoDBP5, showed decreased conidiation and pathogenicity, respectively. We also characterized two transformants that T-DNA was inserted in functionally redundant genes encoding alpha-glucosidase and alpha-mannosidase. These transformants also showed decreased mycelial growth and pathogenicity, implying successful application of this platform in functional analysis of the genes. Our data also demonstrated that comparative phenotypic analysis under over-expression and suppression of gene expression could prove a highly efficient system for functional analysis of the genes. Our over-expressed transformants library would be a valuable resource for functional characterization of the redundant or lethal genes in M. oryzae and this system may be applicable in other fungi.

• 한국유기농업학회지
• /
• 제30권3호
• /
• pp.393-407
• /
• 2022

본 논문에서는 Neobacillus sp. JC05의 추출물을 처리한 오이 식물체에서 고구마뿌리혹선충 감염에 대한 식물의 방어 반응을 조사하였다. Neobacillus sp. JC05의 추출물을 처리한 오이 식물체에서는 고구마뿌리혹선충에 의한 감염이 감소하여 control에 비하여 병 발생이 감소하였으며, 식물이 받는 피해도 줄어들었다. 이때 Neobacillus sp. JC05 추출물을 처리한 오이 식물체의 병 저항성 유전자의 발현량을 관찰한 결과, chitinase와 lipoxygenase 유전자의 발현량이 상대적으로 증가하였으며, 식물체 내 산화 스트레스를 감소시키는 항산화효소 중 peroxidase의 활성이 현저히 증가하는 것을 관찰하였다. 또한, Neobacillus sp. JC05 추출물을 처리한 오이 식물체의 토양 FDase 활성이 증가하였으며, 반면 인산가용화효소와 유레아제 활성은 control에 비해 유의하지 않았다. 그러므로, 이전의 연구 결과와 종합하면, Neobacillus sp. JC05의 추출물은 직접적으로 고구마뿌리혹선충의 알 부화율을 억제하고 유충의 사멸률을 높여 토양 내 고구마뿌리혹선충의 밀도를 감소시키기도 하지만 본 연구의 결과에 따르면, 오이 식물체의 고구마뿌리혹선충에 대한 방어 반응을 증가시켜 저항성을 유도하며, 이를 통해 고구마뿌리혹선충병 발생을 감소시키는데 기여하는 것으로 사료된다.

• 대한수의학회지
• /
• 제46권4호
• /
• pp.355-361
• /
• 2006

Porcine epidemic diarrhea virus (PEDV) strain Chinju99, which was previously isolated from piglets suffering from severe diarrhea was used to characterize the membrane (M) protein gene to establish the molecular information, and the results will be useful in elucidating concepts related to molecular pathogenesis and antigenic structures of PEDV isolates. The Chinju99 M gene generated by reverse transcription and polymerase chain reaction (RT-PCR) consisted of 681 bases containing 22.3% adenine, 22.3% cytosine, 23.1% guanine and 32.3% thymine nucleotides, and the GC content was 45.4%. It had some nucleotide mismatches from M gene of other PEDV strains, such as CV777, Br1/87, KPEDV-9, JMe2, JS2004-2 and LJB-03 with 97-99% nucleotide sequence homology to these strains. Also, it encoded a protein of 226 amino acids, which had some mismatches from those of CV777, Br1/87, KPEDV-9, JMe2, JS20004-2 and LJB-03, as the amino acid sequence homology showed a 97-98% to these strains. The Chinju99 had a very close relationship to the Japanese strain JMe2 for the nucleotide and amino acid sequences of the M gene. The amino acids predicted from Chinju99 M gene consisted of mostly hydrophobic residues and contained three potential sites for asparagine (N)-linked glycosylation, two serine (S)-linked phosphorylation sites by protein kinase C, and two S- or threonine (T)-linked phosphorylation sites by casein kinase II.

• Animal cells and systems
• /
• 제9권4호
• /
• pp.239-244
• /
• 2005

A cDNA clone for a wound- and pathogen-induced gene in Chinese cabbage (Brassica rapa subsp. pekinensis) was isolated and characterized. The cabbage gene, designated BrPR4, encodes a pathogenesis-related protein 4 (PR4) of 140 amino acids. The BrPR4 protein shows high similarity with wound-inducible antifungal proteins of tobacco, potato, barley, and wheat. The BrPR4 gene is locally induced by a nonhost pathogen, Pseudomonas syringae pv. tomato, that elicits a hypersensitive response in Chinese cabbage. Treatment of the cabbage leaves with benzothiadiazole (BTH), methyl jasmonate or ethephon showed that the BrPR4 gene expression is strongly induced by ethylene, but not by methyl jasmonate or BTH. The BrPR4 gene is also activated by wounding. Interestingly, however, the wound-inducible BrPR4 gene expression is repressed by salicylic acid or BTH, suggesting that there is cross-talk between salicylate-dependent and -independent signaling pathways.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권8호
• /
• pp.4509-4513
• /
• 2013

Chronic hepatitis B virus (HBV) infection has long been the most common cause of hepatocellular carcinoma (HCC). However, some aspects of the pathogenesis of HBV infection and genesis of hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC) are still inconclusive. An increasing number of published studies indicate that hepatitis B virus mutations are associated with risk of HCC. These variations include, in particular, mutations in ORF S,C,X gene regions. This mini-review summarizes results of clinical studies and molecular mechanisms on the possible relations of HBV mutations with the development of hepatocellular carcinoma.

• Genomics & Informatics
• /
• 제17권1호
• /
• pp.2.1-2.12
• /
• 2019

Chronic obstructive pulmonary disease (COPD) is a type of progressive lung disease, featured by airflow obstruction. Recently, a comprehensive analysis of the transcriptome in lung tissue of COPD patients was performed, but the heterogeneity of the sample was not seriously considered in characterizing the mechanistic dysregulation of COPD. Here, we established a new transcriptome analysis pipeline using a deconvolution process to reduce the heterogeneity and clearly identified that these transcriptome data originated from the mild or moderate stage of COPD patients. Differentially expressed or co-expressed genes in the protein interaction subnetworks were linked with mitochondrial dysfunction and the immune response, as expected. Computational protein localization prediction revealed that 19 proteins showing changes in subcellular localization were mostly related to mitochondria, suggesting that mislocalization of mitochondria-targeting proteins plays an important role in COPD pathology. Our extensive evaluation of COPD transcriptome data could provide guidelines for analyzing heterogeneous gene expression profiles and classifying potential candidate genes that are responsible for the pathogenesis of COPD.

• BMB Reports
• /
• 제43권10호
• /
• pp.704-709
• /
• 2010

The Swedish mutation (K595N/M596L) of amyloid precursor protein (APP-swe) has been known to increase abnormal cleavage of cellular APP by Beta-secretase (BACE), which causes tau protein hyperphosphorylation and early-onset Alzheimer's disease (AD). Here, we analyzed the effect of APP-swe in global gene expression using deep transcriptome sequencing technique. We found 283 genes were down-regulated and 348 genes were up-regulated in APP-swe expressing H4-swe cells compared to H4 wild-type cells from a total of approximately 74 million reads of 38 base pairs from each transcriptome. Two independent mechanisms such as kinase and phosphatase signaling cascades leading hyperphosphorylation of tau protein were regulated by the expression of APP-swe. Expressions of catalytic subunit as well as several regulatory subunits of protein phosphatases 2A were decreased. In contrast, expressions of tau-phosphorylating glycogen synthase kinase $3\beta$(GSK-3$\beta$), cyclin dependent kinase 5 (CDK5), and cAMP-dependent protein kinase A (PKA) catalytic subunit were increased. Moreover, the expression of AD-related Aquaporin 1 and presenilin 2 expression was regulated by APP-swe. Taken together, we propose that the expression of APP-swe modulates global gene expression directed to AD pathogenesis.

• 한국동물학회:학술대회논문집
• /
• 한국동물학회 2000년도 한국생물과학협회 학술발표대회
• /
• pp.269.2-269
• /
• 2000

No Abstract, See Full Text