• Asian-Australasian Journal of Animal Sciences
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• 제23권1호
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• pp.7-12
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• 2010

The full-length cDNA of the porcine peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase (PECI) gene encodes a monofunctional peroxisomal ${\Delta}^3$,${\Delta}^2$-enoyl-CoA isomerase. Cloning and sequencing of the porcine PECI cDNA revealed the presence of an 1185-base pair open reading frame predicted to encode a 394-amino acid protein by the 5'rapid amplification of cDNA ends (5'RACE) and EST sequences. The porcine PECI gene was expressed in seven tissues (heart, liver, spleen, lung, kidney, skeletal muscle, fat) which was revealed by reverse transcriptase-polymerase chain reaction (RT-PCR). The porcine PECI was mapped to SSC71/2 p11-13 using the somatic cell hybrid panel (SCHP) and the radiation hybrid panel (RH) (LOD score 12.84). The data showed that PECI was closely linked to marker S0383. A C/T single nucleotide polymorphism in PECI exon 10 (3'UTR) was detected as a PvuII PCR-RFLP. Association analysis in our experimental pig population showed that different genotypes of PECI gene were significantly associated with the Average Backfat thickness (ABF) (p<0.05) and Buttock backfat thickness (p<0.01).

• Asian-Australasian Journal of Animal Sciences
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• 제19권7호
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• pp.931-937
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• 2006

The full-length cDNA of the porcine SMPX gene was obtained by the rapid amplification of cDNA ends (RACE). The nucleotide sequences and the predicted protein sequences share high sequence identity with both human and mouse. The promoter of SMPX was sequenced and then analyzed to find the promoter binding sites. The reverse transcriptase-polymerase chain reaction (RT-PCR) revealed that SMPX has a high level of expression in heart and skeletal muscle, a very low expression in lung and spleen and no expression in liver, kidney, fat and brain. Moreover, SMPX has a differential expression level in skeletal muscle, the expression in 65-day embryos being higher than other stages. The porcine SMPX was mapped to SSCXp24 by using a somatic cell hybrid panel (SCHP) and was found closely linked to SW1903 using the radiation hybrid panel IMpRH. An A/G single nucleotide polymorphism (PCR-RFLP) in the 3'-untranslated region (3'-UTR) was detected in eight breeds. The analysis of allele frequency distribution showed that introduced pig breeds (Duroc and Large White) have a higher frequency of allele A while in the Chinese indigenous pig breeds (Qingping pig, Lantang pig, YushanBlack pig, Large Black-White pig, Small Meishan) have a higher frequencies of allele G. The association analysis using an experimental population (188 pigs), which included two cross-bred groups and three pure-blood groups, suggested that the SNP genotype was associated with intramuscular fat content.

• Journal of Microbiology and Biotechnology
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• 제12권5호
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• pp.711-715
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• 2002

Telomerase adds telomeric repeats to chromosomal ends and is known to play an important role in carcinogenesis through cellular immortalization. Since telomerase is an essential pathogenomic factor in malignant tumors, inhibiting telomerase activity is thought to be possible to make telomerase positive tumors more sensitive to cisplatin treatment, which is effective in ovarian cancers, but clinical success Is limited by chemo-resistance. In the present study, cisplatin-sensitive ovarian cancer cell line A2780 and cisplatin-resistant A2780/cp70 cell line were infected with antisense telomerase adenovirus Ad-OA. It was found that the Ad-OA suppressed ovarian cancer cell growth and this effect was mainly due to the induction of caspase-dependent apoptosis. Next, we infected the cisplatin resistant ovarian cancer cell line A2780/ cp70 with Ad-OA and cisplatin concurrently. Interestingly, cisplatin treatment with Ad-Oh was more effective to cisplatin-induced cell death in A2780/cp70 cells compared to cisplatin or the vector group only. These data suggest that cisplatin treatment with Ad-OA may be a new chemo-sensitizer for cisplatin resistant ovarian cancer.

• 미생물학회지
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• 제38권3호
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• pp.221-229
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• 2002

일본뇌염바이러스(Japanese encephalitis virus, JEV) Nakayama strain을 초기 multiplicity of infection 5.0으로 Vero세포에 감염하여 1년 이상 안정적으로 바이러스를 방출하는 지속감염(persistently-infected)세포주를 확립하였다. 지속감염 세포에서 지속적으로 방출되는 총 11 개의 Plaque 형태 변이바이러스(morphology mutants)클론을 확보하였다. 분리된 변이바이러스의 복제효율을 분석한 결과 생물학적 표현형과 복제효율은 유의하게 상관하였다. 변이바이러스 RNA 게놈 양 말단의 non-coding region 및 envelop 단백질의 ORF에서는 유의한 염기서열 변화가 관찰되지 않아 JEV 약독화에 새로운 인자가 추가로 관여할 가능성을 제시하였다. 변이바이러스에 감염된 신선한 Vero세포는 wild-type JEV의 일반적 감염성상과 다르게 대다수의 세포가 유의할 만한 세포병변현상을 나타내지 않았다. 감염된 Vero세포에서 wild-type JEV 및 large plaque을 형성하는 변이바이러스의 경우 mRNA와 함께 Bcl-2의 발현은 모두 유의하게 감소하였으며 p53은 뚜렷하게 증가하였다. 반면 small plaque을 형성하는 변이바이러스의 감염세포에서는 Bcl-2와 p53 모두 유의한 변화를 볼 수 없었다. 이상의 결과들과 함께, 감염된 Vero세포의 internucleosomal DNA fragmentation과DNA profile의 유형분석에 따르면 궁극적 인 세포병변효과의 변화는 변이바이러스의 복제효율과 더불어 p53에 비의존적인 apoptosis 수위의 전반적인 감소에 기인하는 것으로 판단된다.

• Nutrition Research and Practice
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• 제13권1호
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• pp.58-63
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• 2019

BACKGROUND/OBJECTIVES: Telomeres are located at the chromosomal ends and progressively shortened during each cell cycle. Telomerase, which is regulated by hTERT and c-MYC, maintains telomeric DNA sequences. Especially, telomerase is active in cancer and stem cells to maintain telomere length for replicative immortality. Recently we reported that walnut phenolic extract (WPE) can reduce cell viability in a colon cancer stem cell (CSC) model. We, therefore, investigated the effect of WPE on telomere maintenance in the same model. MATERIALS AND METHODS: $CD133^+CD44^+$ cells from HCT116, a human colon cancer cell line, were sorted by Fluorescence-activated cell sorting (FACS) and treated with WPE at the concentrations of 0, 10, 20, and $40{\mu}g/mL$ for 6 days. Telomere lengths were assessed by quantitative real-time PCR (qRT-PCR) using telomere specific primers and DNA extracted from the cells, which was further adjusted with single-copy gene and reference DNA ($ddC_t$). Telomerase activity was also measured by qRT-PCR after incubating the PCR mixture with cell protein extracts, which was adjusted with reference DNA ($dC_t$). Transcriptions of hTERT and c-MYC were determined using conventional RT-PCR. RESULTS: Telomere length of WPE-treated cells was significantly decreased in a dose-dependent manner ($5.16{\pm}0.13$ at $0{\mu}g/mL$, $4.79{\pm}0.12$ at $10{\mu}g/mL$, $3.24{\pm}0.08$ at $20{\mu}g/mL$ and $3.99{\pm}0.09$ at $40{\mu}g/mL$; P = 0.0276). Telomerase activities concurrently decreased with telomere length ($1.47{\pm}0.04$, $1.09{\pm}0.01$, $0.76{\pm}0.08$, and $0.88{\pm}0.06$; P = 0.0067). There was a positive correlation between telomere length and telomerase activity (r = 0.9090; P < 0.0001). Transcriptions of both hTERT and c-MYC were also significantly decreased in the same manner. CONCLUSION: In the present cell culture model, WPE reduced telomere maintenance, which may provide a mechanistic link to the effect of walnuts on the viability of colon CSCs.