• Asian-Australasian Journal of Animal Sciences
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• 제21권4호
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• pp.499-509
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• 2008

Foxo1 plays an important role in the integration of hormone-activated signaling pathways with the complex transcriptional cascade that promotes preadipocyte differentiation of clonal cell lines from rodents. We isolated the full-length cDNA of porcine FoxO1 gene using RACE, confirmed by visual Northern blotting. The deduced amino acids indicated 94% and 90% identities with the corresponding human and mice aa. Analysis of the aa sequence, showed that it included a Forkhead domain (aa 167-247), a transmembrane structure domain (aa 90-113), a LXXLL motif (aa 469-473), and 51 Ser, 8 Thr, and 4 Tyr phosphorylation sites, indicating a potential important role for FoxO1 transcriptional activity in vivo. Using the IMpRH panel, we mapped FoxO1 gene to chromosome 11p13. Our data provide basic molecular information useful for the further investigation on the function of FoxO1 gene. Time-course analysis of FoxO1 expressions indicated that levels of mRNA and protein gradually increased from day 0 to 3, and it reached almost maximal level at day 3, then decreased from day 5 to 7 in porcine primary preadipocyte differentiation. After induction by IGF-1, GPDH activity and accumulation of lipid increased, however, expressions of FoxO1 mRNA and protein were inhibited in a dose dependent manner. These results suggest that FoxO1 takes part in porcine preadipocyte differentiation and expressions of FoxO1 were regulated by IGF-1.

• Journal of Microbiology and Biotechnology
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• 제18권5호
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• pp.828-836
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• 2008

The biocontrol rhizobacterium Pseudomonas sp. M18 can produce two kinds of antibiotics, namely pyoluteorin (Plt) and phenazine-1-carboxylic acid (PCA), and is antagonistic against a number of soilborne phytopathogens. In this study, a luxR-type quorum-sensing regulatory gene, vqsR, was identified and characterized immediately downstream of the Plt gene cluster in strain MI8. A vqsR-inactivated mutant led to a significant decrease in the production of Plt and its biosynthetic gene expression. However, this was restored when introducing the vqsR gene by cloning into the plasmid pME6032 in trans. The vqsR mutation did not exert any obvious influence on the production of PCA and its biosynthetic gene expression and the production of N-acylhomoserine lactones (C4 and C8-HSLs) and their biosynthetic gene rhlI expression. Accordingly, these results introduce VqsR as a regulator of Plt production in Pseudomonas spp., and suggest that the regulatory mechanism of vqsR in strain M18 is distinct from that in P. aeruginosa. In addition, it was demonstrated that vqsR mutation did not have any obvious impact on the expression of Plt-specific ABC transporters and other secondary metabolic global regulators, including GacA, RpoS, and RsmA.

• 한국환경성돌연변이발암원학회지
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• 제19권1호
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• pp.28-33
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• 1999

The SNF2 family includes proteins from a variety of species with roles I cellular processes such as transcriptional regulation, recombination and various types of DNA repair. Several proteins with unknown function are also included in this family. Here, we report the cloning and characterization of hrp 2+ gene (helicase related gene from S. pombe) which was isolated by PCR amplication using the conserved domain of SNF2 motifs within the ERCC6 gene which encodes a protein involved in DNA excision repair. The hrp2+ gene was isolated by screening with yeast S. pombe genomic library. The isolated cloned contained 6.5 kb insert DNA. Southern blot analysis confirmed that S. pombe chromosome contains the same DNA as hrp2+ gene and this gene exists as a single copy in S. pombe genome. The 4.7 kb transcript of mRNA was identified by Northern blot. To examined the transcriptional regulation of hrp2+ gene, DNA damaging agents were treated. These results indicated that the hrp2+ gene may not be directly involved in DNA replication, but may be involved in damage response pathway.

• Journal of Plant Biotechnology
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• 제34권3호
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• pp.173-180
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• 2007

The full-length cDNA of LeAPR1 encoded a protein of 461 amino acid residues, which contained homology with phosphoadenosine phosphosulphate reductase (PAPS reductase) in N-terminal and an adenylylsulfate reductase in N-term and C-terminal. Analysis of the deduced amino acid sequence of LeAPR1 revealed that it shares high sequence identity with potato StAPR (96% identity)(Gene bank accession no. CDC44841). We found that multiple copies of LeAPR1 gene are present in the tomato genome through southern blot using genomic DNA was digested with 3 different restriction enzymes. The expression of LeAPR1 was also examined in various organs and its expression was also detected at high levels in roots and stems. Only high amounts of LeAPR1 transcripts were detected at high transcripts in the leaves at time 0, and then reduced as the plant stressed by the NaCl and abscisic acid (ABA). After 24h treatment of NaCl and ABA were showed increasing patterns of LeAPR1 gene. Time course of LeAPR1 gene expression was examined under oxidative stresses from metyl viologen (MV) and hydrogen peroxide ($H_2O_2$). In the presence of 10 mM $H_2O_2$ and $50\;{\mu}M$ MV, the levels of LeAPR1 transcript in leaves decreased after 1 h, and then increased strongly, peaked at 24 h. Our results indicated that LeAPR1 may play a role function of circadian regulation involved in abiotic stresses signaling pathways.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2002년도 창립10주년기념 및 국립독성연구원 의약품동등성평가부서 신설기념 국재학술대회:생물학적 동등성과 의약품 개발 전략을 위한 국제심포지움
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• pp.238-238
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• 2002

We studied whether kinase pathways are involved in TCDD-induced gene expression by treating specific kinase inhibitors ncluding MEK1 inhibitor PD98059, p38 inhibitor SB202190, PI-3 kinase inhibitor Wortmannin or LY294002 or protein tyrosine kinase inhibitor Genestein and then tested the effects of individual inhibitors on TCDD-induced gene expression of cytochromelAl gene (CYPlAl). Our results show that PD98059, MEK-1 inhibitor reduces dioxin-inducible transcription of CYPlAl. p44/p42MAPK, that is phosphorylated by Mek-1, are phosphorlylated by treatment of TCDD, peaking at lnM, 30min treatments. Overexpressions of p44/p42 MAPK dominant negative mutants suppress dioxin dependent transcription of DRE-driven reporter gene in a dose-dependent manner. Our results demonstrate that p44/p42 MAPK is essential for transcriptional activity of AHR/ARNT heterodimer. We found that PD98059 dose-dependently blocks TCDD-induced DRE binding of the AHR/ARNT heterodimer, thereby it reduces TCDD-induced gene expression. Therefore, our results indicate that Mek-1/p44/p42 MAPK pathway is involved in TCDD-induced gene expression, [This study was supported by a grant from Korean Research Foundation Grant (X01529)to H. Park]

• 한국작물학회지
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• 제49권2호
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• pp.148-153
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• 2004

The Hessian fly, Mayetiola destructor (Say), is known to be one of the major insect herbivores of wheat worldwide. In order to provide molecular events on interactions of the NIL with H21 and larvae of Hessian fly biotype L, the TaCR1 gene, Triticum aestivum cytokinin repressed 1, was isolated through the suppression subtractive hybridization, which was constructed using stems of the NIL with H21 at 6 days after infestation as tester and stems of the recurrent parent Coker797 without H21 at 6 days after infestation as driver. Transcript levels of TaCR1 mRNA in the NIL with H21 were highest at 6 days after infestation but in the Coker797 without H21 until 8 days were similar with those of non-infested plants. Expression of the TaCR1 gene was decreased at early time and then recovered after wounding or $H_2O$$_2$ treatment as well as 6-BAP treatment. Transcripts levels of the TaCR1 gene was changed after MeJA, SA, ethephone, or ABA treatment. In drought treatment, the TaCRl gene were increased at early stage of stress and then decreased at late stage. Expression of the TaCRl gene was continued to decrease through 24 h in the cold treatment. Although the TaCRl gene is increased through infestation in NIL with H21, further study was required to elucidate a role on resistance against larvae of Hessian fly. However, the TaCR1 gene could be used as marker gene on response of plants against abiotic stresses as well as application of plants with several hormones.

• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.4149-4154
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• 2016

Background: Cervical carcinoma is one of the main causes of mortality in women worldwide as well as in India. It occurs as a result of various molecular events that develop from the combined influences of an individual's genetic predisposition and external agents such as smoking and menstrual hygiene, for example. However, infection with human papillomavirus (HPV) is the established major risk factor. The aim of the current study was to investigate p16 CpG island methylation and establish any correlation with mRNA expression in north Indian population. Materials and Methods: We analyzed 196 woman volunteer out of which 98 were cases and 98 healthy controls. For the analysis of methylation pattern, DNA extracted from blood samples was modified with a bisulfate kit and used as template for methylation specific PCR (MSP). Quantitative real-time PCR (QRT-PCR) was performed to check mRNA expression. Results: Correlation between methylation status of p16 gene and poor menstrual hygiene was significant (p=0.006), high parity cases showed methylation of p16 gene (p=0.031) with increased risk up to 1.86 times for cervical cancer and smoking was a strong risk factor associated with cervical cancer. We analyzed methylation pattern and found 60.3% methylation in cases with low mRNA expression level (0.014) as compare to controls (1.24). It was also observed that promoter methylation of p16 gene was significantly greater in FIGO stage III. Conclusions: We conclude that p16 methylation plays an important role in cervical cancer in the north Indian population and its methylation decreases mRNA expression. It can be used as an important and consistent blood biomarker in cervical cancer patients.

• 한국축산식품학회지
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• 제37권2호
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• pp.168-174
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• 2017

We hypothesized that supplementing finishing diets with palm oil would promote adipogenic gene expression but depress stearoyl-CoA desaturase (SCD) gene expression in intramuscular (i.m.) adipose tissues of Hanwoo steers during fattening period (from 16 to 32 mon of age). Fourteen Hanwoo steers were allotted randomly to 2 groups of 7 steers based on initial BW and fed either a basal diet (control) or the basal diet supplemented with 5% palm oil (BDSP). At slaughter, i.m. adipose tissue was harvested for analysis of adipogenic gene expression and fatty acid composition. There were no differences in BW or average daily gain between treatment groups. Supplemental palm oil had no effect on carcass quality traits (carcass weight, backfat thickness, loin muscle area, or marbling scores) or meat color values. Palm oil increased (p<0.05) expression of AMP-activated protein kinase-${\alpha}$ and peroxisome proliferator-activated receptor-${\gamma}$, but decreased (p<0.05) CAAT/enhancer binding protein-${\beta}$ gene expression and tended to decrease stearoyl-CoA desaturase gene expression in i.m. adipose tissue. Palm oil increased total i.m. polyunsaturated fatty acids (p<0.05) compared to the control i.m. adipose tissue, but had no effect on saturated or monounsaturated fatty acids. Although there were significant effects of supplemental palm oil on i.m. adipose tissue gene expression, the absence of negative effects on carcass and meat characteristics indicates that palm oil could be a suitable dietary supplement for the production of Hanwoo beef cattle.

• Journal of Microbiology and Biotechnology
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• 제12권1호
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• pp.114-120
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• 2002

A 1,989-bp genomic region encoding nickel resistance genes was isolated from Legionella pneumophila, a pathogen for legionellosis. From a sequencing and computer analysis, the region was found to harbor two structural genes, a nreB-like protein gene (1,149 bp) and a nreA-like protein gene (270 bp), in a row. Both genes exhibited a significant degree of similarity to the corresponding genes from Synechocystis sp. PCC6803 ($54\%$ amino acid sequence identity) and Achromobacter xylosoxidans 31A ($76\%$). The gene was successfully expressed in E. coli MG1655 and conferred a nickel resistance of up to 5 mM in an LB medium and 3 mM in a TMS medium including gluconate as the sole carbon source. E. coli harboring the nickel resistance gene also exhibited a substantial resistance to cobalt, yet no resistance to cadmium or zinc. Since the extracellular concentration of nickel remained constant during the whole period of cultivation, it was confirmed that the nickel resistance was provided by an efflux system like the $Ni^2＋$permease (nrsD) of Synechocystis sp. strain PCC6803. Since polyphosphate (poly-P) is known as a global regulator for gene expression as well as a potential virulence factor in E. coli, the nickel resistance of a ppk mutant of E. coli MG 1655 harboring the nickel resistance gene from L. pneumophila was compared with that of its parental strain. The nickel resistance was significantly attenuated by ppk inactivation, which was more pronounced in an LB medium than in a TMS medium.

• Reproductive and Developmental Biology
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• 제35권4호
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• pp.427-434
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• 2011

Mitochondria diseases have been reported to involve structural and functional defects of complex I-V. Especially, many of these diseases are known to be related to dysfunction of mitochondrial proton-translocating NADH-ubiquinone oxidoreductase (complex I). The dysfunction of mitochondria complex I is associated with neurodegenerative disorders, such as Parkinson's disease, Huntington's disease, and Leber's hereditary optic neuropathy (LHON). Mammalian mitochondrial proton-translocating NADH-quinone oxidoreductase (complex I) is largest and consists of at least 46 different subunits. In contrast, the NDI1 gene of Saccharomyces cerevisiae is a single subunit rotenone-insensitive NADH-quinone oxidoreductase that is located on the matrix side of the inner mitochondrial membrane. The Saccharomyces cerevisiae NDI1 gene using a recombinant adeno-associated virus vector (rAAV-NDI1) was successfully expressed in AML12 mouse liver hepatocytes and the NDI1-transduced cells were able to grow in media containing rotenone. In contrast, control cells that did not receive the NDI1 gene failed to survive. The expressed Ndi1 enzyme was recognized to be localized in mitochondria by confocal immunofluorescence microscopic analyses and immunoblotting. Using digitonin-permeabilized cells, it was shown that the NADH oxidase activity of the NDI1-transduced cells was not affected by rotenone which is inhibitor of complex I, but was inhibited by antimycin A. Furthermore, these results indicate that Ndi1 can be functionally expressed in the AML12 mouse liver hepatocytes. It is conceivable that the NDI1 gene is powerful tool for gene therapy of mitochondrial diseases caused by complex I deficiency. In the future, we will attempt to functionally express the NDI1 gene in mouse embryonic stem (mES) cell.