• 한국버섯학회지
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• 제18권1호
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• pp.100-106
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• 2020

본 연구는 국내외 수집 균주를 대상으로 균주의 배양적 특성과 유전적 유연관계 및 균주별 함유된 베타글루칸의 함량을 분석하였다. 곰보버섯은 생장온도 25℃, pH 7.0에서 균사 생장이 가장 왕성하였다. 균사는 초기에 백색에서 생장이 진행될수록 진한 노란색을 띠다 진한 갈색으로 변화하는 공통적인 특징을 가지고 있었으며, 곰보버섯만의 고유한 배양적 특징으로 균사가 종에 따라 주기별로 경화되는 특징과 특유 강한 향이 있다는 것을 확인할 수 있었다. 균사 형태는 관찰을 통하여 총 5종류로 분류되었고 이들 균주의 ITS 분석 결과 Morchella conica, M.sextelata, M. importuna, M. esculenta, M. carssipes 등 5종으로 동정되었다. UFPF primer를 이용하여 PCR 다형성을 분석한 결과 ITS 분석 결과와는 다르게 M. conica로 동정된 KMCC04971 균주와 M. sextelata로 동정된 KMCC04407 균주는 동일한 패턴을 보였으며 그 결과 4개의 그룹으로 분류할 수 있었다. 균주별 균사체 베타글루칸 함량 분석 결과 M. importuna인 KMCC04973 균주가 100 g당 알파글루칸 16.4 g을 포함하는 베타글루칸 함량 33.1 g으로 가장 높았다.

• Asian Pacific Journal of Cancer Prevention
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• 제17권8호
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• pp.3971-3977
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• 2016

Background: Breast cancer, the commonest cancer among women in the world, ranks top in India with an incidence rate of 1,45,000 new cases and mortality rate of 70,000 women every year. Chemotherapy outcome for breast cancer is hampered due to poor response and irreversible dose-dependent cardiotoxicity which is determined by genetic variations in drug metabolizing enzymes and transporters. Pregnane X receptor (PXR), a member of the nuclear receptor superfamily, induces expression of drug metabolizing enzymes (DMEs) and transporters leading to regulation of xenobiotic metabolism. Materials and Methods: A genomic region spanning PXR 3' UTR was amplified and sequenced using genomic DNA isolated from 96 South Indian breast cancer patients. Genetic variants observed in our study subjects were queried in miRSNP to establish SNPs that alter miRNA binding sites in PXR 3' UTR. In addition, enrichment analysis was carried out to understand the network of miRNAs and PXR in drug metabolism using DIANA miRpath and miRwalk pathway prediction tools. Results: In this study, we identified SNPs rs3732359, rs3732360, rs1054190, rs1054191 and rs6438550 in the PXR 3; UTR region. The SNPs rs3732360, rs1054190 and rs1054191 were located in the binding site of miR-500a-3p, miR-532-3p and miR-374a-3p resulting in the altered PXR level due to the deregulation of post-transcriptional control and this leads to poor treatment response and toxicity. Conclusions: Genetic variants identified in PXR 3' UTR and their effects on PXR levels through post-transcriptional regulation provide a genetic basis for interindividual variability in treatment response and toxicity associated with chemotherapy.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6261-6265
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• 2013

Background: Recent studies have suggested that expression of the RAS protein activator like-1 gene (RASAL1) is decreased in gastric carcinoma tissues and cell lines, indicated a role in tumorigenesis and development of gastric cancer. Reduced expression of RASAL1 could result in aberrant increase of activity of RAS signaling pathways in cancer cells. However, the exact mechanism which induces down-regulation of the RASAL1 gene remains unclear. This study aimed to determine the methylation status and regulation of RASAL1 in gastric cancer. Materials and Methods: Using the methylation-specific polymerase chain reaction (MSP), the methylation status of CpG islands in the RASAL1 promoter in gastric cancers and paired adjacent non-cancerous tissues from 40 patients was assessed and its clinicopathological significance was analyzed. The methylation status of RASAL1 in gastric cancer lines MKN-28, SGC-790l, BGC-823, as well as in normal gastric epithelial cell line GES-l was also determined after treatment with a DNA methyltransferase inhibitor, 5-aza-2'-doexycytidine (5-Aza-CdR). RAS activity (GAS-GTP) was assessed through a pull-down method, while protein levels of ERK1/2, a downstream molecule of RAS signaling pathways, were determined by Western blotting. Results: The frequencies of RASAL1 promoter methylation in gastric cancer and paired adjacent non-cancerous tissues were 70% (28/40) and 30% (12/40) respectively (P<0.05). There were significantly correlations between RASAL1 promoter methylation with tumor differentiation, tumor size, invasive depth and lymph node metastasis in patients with gastric cancer (all P<0.05), but no correlation was found for age or gender. Promoter hypermethylation of the RASAL1 gene was detected in MKN-28, SGC-790l and BGC-823 cancer cells, but not in the normal gastric epithelial cell line GES-1. Elevated expression of the RASAL1 protein, a decreased RAS-GTP and p-ERK1/2 protein were detected in three gastric cancer cell lines after treatment with 5-Aza-CdR. Conclusions: Aberrant hypermethylation of the RASAL1 gene promoter frequently occurs in gastric cancer tissues and cells. In addition, the demethylating agent 5-Aza-CdR can reverse the hypermethylation of RASAL1 gene and up-regulate the expression of RASAL1 significantly in gastric cancer cells in vivo. Our study suggests that RASAL1 promoter methylation may have a certain relationship with the reduced RASAL1 expression in gastric cancer.

• Journal of Microbiology and Biotechnology
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• 제18권7호
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• pp.1290-1297
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• 2008

To investigate the diversities of Accumulibacter phosphatis and its polyhydroxyalkanoate (PHA) synthase gene (phaC) in enhanced biological phosphorus removal (EBPR) sludge, an acetate-fed sequencing batch reactor was operated. Analysis of microbial communities using fluorescence in situ hybridization and 16S rRNA gene clone libraries showed that the population of Accumulibacter phosphatis in the EBPR sludge comprised more than 50% of total bacteria, and was clearly divided into two subgroups with about 97.5% sequence identity of the 16S rRNA genes. PAO phaC primers targeting the phaC genes of Accumulibacter phosphatis were designed and applied to retrieve fragments of putative phaC homologs of Accumulibacter phosphatis from EBPR sludge. PAO phaC primers targeting $G_{1PAO},\;G_{2PAO},\;and\;G_{3PAO}$ groups produced PCR amplicons successfully; the resulting sequences of the phaC gene homologs were diverse, and were distantly related to metagenomic phaC sequences of Accumulibacter phosphatis with 75-98% DNA sequence identities. Degenerate NPAO (non-PAO) phaC primers targeting phaC genes of non-Accumulibacter phosphatis bacteria were also designed and applied to the EBPR sludge. Twenty-four phaC homologs retrieved from NPAO phaC primers were different from the phaC gene homologs derived from Accumulibacter phosphatis, which suggests that the PAO phaC primers were specific for the amplification of phaC gene homologs of Accumulibacter phosphatis, and the putative phaC gene homologs by PAO phaC primers were derived from Accumulibacter phosphatis in the EBPR sludge. Among 24 phaC homologs, a phaC homolog (GINPAO-2), which was dominant in the NPAO phaC clone library, showed the strongest signal in slot hybridization and shared approximately 60% nucleotide identity with the $G_{4PAO}$ group of Accumulibacter phosphatis, which suggests that GINPAO-2 might be derived from Accumulibacter phosphatis. In conclusion, analyses of the 16S rRNA and phaC genes showed that Accumulibacter phosphatis might be phylogenetically and metabolically diverse.

• Asian-Australasian Journal of Animal Sciences
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• 제21권7호
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• pp.917-922
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• 2008

A disintegrin-like and metalloprotease (reprolysin type) with thrombospondin type 1 motif (ADAMTS1) plays a critical role in follicular rupture and represents a major advance in the proteolytic events that control ovulation. In this study, a 9,026-bp DNA sequence containing the full coding region, all 8 introns and part of the 5'and 3' untranslated region of the porcine ADAMTS1 gene was obtained. Analysis of the ADAMTS1 gene using the porcine radiation hybrid panel indicated that pig ADAMTS1 is closely linkage with microsatellite marker S0215, located on SSC13q49. The open reading frame of its cDNA covered 2,844 bp and encoded 947 amino acids. The coding region of porcine ADAMTS1 as determined by sequence alignments shared 85% and 81% identity with human and mouse cDNAs, respectively. The deduced protein contained 947 amino acids showing 85% sequence similarity both to the human and mouse proteins, respectively. Comparative sequencing of three pig breeds revealed one single nucleotide polymorphism (SNP) within exon 7 of which a G-C substitution at position 6006 changes a codon for arginine into a codon for proline. The substitution was situated within a PvuII recognition site and developed as a PCR-RFLP marker for further use in population variation investigations and association analysis with litter size. Allele frequencies of this SNP were investigated in seven pig breeds/lines. An association analysis in a new Qingping female line suggested that different ADAMTS1 genotypes have significant differences in litter size (p<0.01).

• Asian Pacific Journal of Cancer Prevention
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• 제16권16호
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• pp.7071-7076
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• 2015

Epidermal growth factor receptor (EGFR) is one of the targeted molecular markers in many cancers including lung malignancies. Gefitinib and erlotinib are two available therapeutics that act as specific inhibitors of tyrosine kinase (TK) domains. We performed a case-control study with formalin-fixed paraffin-embedded tissue blocks (FFPE) from tissue biopsies of 167 non-small cell lung carcinoma (NSCLC) patients and 167 healthy controls. The tissue biopsies were studied for mutations in exons 18-21 of the EGFR gene. This study was performed using PCR followed by DNA sequencing. We identified 63 mutations in 33 men and 30 women. Mutations were detected in exon 19 (delE746-A750, delE746-T751, delL747-E749, delL747-P753, delL747-T751) in 32 patients, exon 20 (S786I, T790M) in 16, and exon 21 (L858R) in 15. No mutations were observed in exon 18. The 63 patients with EFGR mutations were considered for upfront therapy with oral tyrosine kinase inhibitor (TKI) drugs and have responded well to therapy over the last 15 months. The control patients had no mutations in any of the exons studied. The advent of EGFR TKI therapy has provided a powerful new treatment modality for patients diagnosed with NSCLC. The study emphasizes the frequency of EGFR mutations in NSCLC patients and its role as an important predictive marker for response to oral TKI in the south Indian population.

• Journal of Crop Science and Biotechnology
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• 제11권2호
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• pp.135-140
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• 2008

Improving eating quality is one of the most important objectives in japonica rice breeding programs in Yunnan Province of China. Eating quality and its relevant traits of nine Korean and 11 Yunnan rice cultivars were comparatively analyzed in this study. The grain shape of most Yunnan japonica rice cultivars have a relatively slender shape and are slightly larger than Korean rice cultivars. Palatability value of cooked rice of Yunnan rice cultivars was significantly lower, while the protein content of Yunnan rice cultivars was significantly higher than that of Korean cultivars. Peak viscosity and breakdown viscosity of the Yunnan rice cultivars were significantly lower, while setback viscosity of the Yunnan rice cultivars was significantly higher than in Korean rice cultivars. Palatability value of cooked rice was negatively correlated with protein content and setback viscosity but positively correlated with peak viscosity, breakdown viscosity, and cool paste viscosity. Through multiple linear regression analysis, an equation for estimating palatability value(PV) of cooked rice based on quality traits was generated as dependent only upon protein content(PC), PV=139.024-(10.865$\times$PC) with an $R^2$ value of 0.822. The results suggest that reducing protein contents should be the major target in improving eating quality of Yunnan japonica rice cultivars through integrated approaches of both cultivar development and appropriate cultural practices. Genetic similarities among cultivars based on DNA markers which had been identified as associated with grain quality seemed not to be directly related to PV.

• 생명과학회지
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• 제19권2호
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• pp.163-168
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• 2009

천초근은 전통적으로 동양의학에서 항암제로 사용되어왔는데 인간 급성 백혈병 세포주인 Jurkat T 세포를 사용하여 천초근의 세포독성기작을 알아보았다. 천초근 뿌리(3 kg)를 메탄올로 추출, 증류한 후 물에 녹여 다시 dichloromethane으로 추출분획 하였다. 세포독성 활성을 보이는 dichloromethane 추출물을 연속적으로 HPLC를 통해 분리하였고 그 활성물질(65 mg)을 CCH1이라 명명하였다. CCH1을 0.5 ${\mu}g$/ml에서 2.0 ${\mu}g$/ml의 농도로 처리하고 세포사멸 과정을 보았다. 즉, mitochondria cytochrome c 방출, casapase-8, -9 및 caspase-3의 활성화, PARP 분해, DNA 단편화 현상들이 일어나는 것을 관찰하였다. 하지만, mitochondria cytochrome c 방출 억제자인 Bcl-xL이 과발현되는 Jurkat T 세포에서는 세포사멸현상이 일어나지 않았다. 이러한 결과는 CCH1이 mitochondria 의존적인 신호전달 과정을 통해서 세포사멸을 유도 한다고 할 수 있다. 그리고 CCH1에 의한 세포독성은 혈액에서 분리한 단핵구 세포보다 Jurkat T 세포에서 보다 강한 활성을 보였다.

• 미생물학회지
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• 제40권2호
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• pp.121-126
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• 2004

4-chlorobiphenyl (4CB)의 분해균주인 Pseudomonas sp. strain DJ-12의 16S rDNA의 염기서열을 분석한 결과 Comamonas sp. strain DJ-12로 재분류 되었다. Pseudomonas sp. strain DJ-12로부터 4CB의 분해결과 생성되는 2,3-dihydroxybiphenyl을 계속 분해하는데 관여하는 pcbC1Dl 유전자를 이미 보고된 바 있다. 이번 연구에서는 Comamonas sp. strain DJ-12로부터 4CB 분해에 관여하는 pcbABC2D2 유전자를 클로닝하여 염기서열을 분석하였다. PcbAB 및 pcbCD 유전자들의 염기서열은 48, 65％, 추정 아미노산 서열은 33, 42％의 낮은 유사도를 보였다. 본 연구에서 얻어진 pcbC2D2 유전자는 이미 보고만 pcbCIDl 유전자와 염기의 개수와 서열의 유사도가 서로 다름을 보여 주었다. Comamonas sp. strain DJ-12의 두 가지 pcbCD유전자들은 Southern hybridization 결과에서도 유사성을 보이지 않았으며, 서로 다른 위치에 존재함을 보여주었다. 그러나 2,3-dihydroxybiphenyl의 분해 특성은 동일하였다. 이와 같은 결과는 Comamonas sp. strain DJ-12 균주가 2조의 pcbCD 유전자를 가지고 있다는 것을 의미하는 것이다.

• 한국식품위생안전성학회지
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• 제32권3호
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• pp.199-205
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• 2017

인수공통 감염증의 하나인 톡소포자충의 검출을 위해서는 대부분은 ELISA 법이 사용 되고 있으나, 충체가 사멸된 후에도 양성반응이 나타나는 등 사용에 제한이 있다. 반면 유전자 검출법은 현재 감염상태를 확인 할 수 있기 때문에 식중독 원인조사 등에 적합하다고 판단되어 이를 활용하여 본 연구를 진행하였다. 톡소포자충의 유전정보를 통해 529 repeat region의 염기서열을 얻고, 프라이머 및 TaqMan 프로브를 설계하여 real-time PCR을 이용한 검출법을 개발하였다. 검출한계(lower limit of detection) 및 적정곡선을 확인한 결과 10 genomic DNA copy가 검출한계로 확인되었고, 정량을 위한 곡선은 $10^1{\sim}10^6$ DNA copies까지 0.999의 $R^2$ 값을 나타내었다. 개발된 검출법의 증폭효율을 비교하기 위해 B1 gene 타겟 프라이머 세트와 타입별 검출한계를 비교한 결과, type 1, 2, 3 톡소포자충에서 같거나 더 나은 검출한계를 보였다. 또한 식품에서 주로 분리되는 식중독 세균 14종 및 원충 3종에 대해 특이도를 비교한 결과, 모두 음성으로 나타났다. 개발된 검출법을 식육검체에 적용하였을 때 type 1, 2, 3에서 모두 원활한 검출결과를 보여 증폭방해물질이 존재하지 않은 것으로 확인되었다. 본 연구를 통해 개발된 유전자검출법은 국내 유통 중인 식육에서 인수 공통감염 원충의 하나인 톡소포자충의 감염 여부를 확인하는 사전적 모니터링의 방법으로 활용될 예정이다.