• Journal of Animal Science and Technology
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• 제51권4호
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• pp.289-294
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• 2009

본 연구는 한우 inositol 1,4,5-triphosphate receptor type1(IP3R1) 유전자를 대상으로 SNP를 발굴하고, 도체형질과의 관련성 분석을 위하여 수행하였다. PCR 및 염기서열 결정법을 통해 IP3R1 유전자내 3개의 SNP를 발굴하였고, 이중 intron 29에 위치하는 SNP의 경우 미 보고된 신규 SNP로 확인되었다. 발굴된 3개의 SNP를 대상으로 표현형 기록치를 보유한 후대검정우 583두에 대하여 유전자형 분석 및 관련성 분석을 수행하였다. 분석결과 3개의 SNP 중 g.1428617A>G SNP가 생시체중(P<0.05) 및 도체중(P<0.01)과 유의적인 상관관계가 있음을 확인할 수 있었다. 이러한 결과는 추후 한우 개량을 위한 유전자 마커로 활용이 가능할 것으로 판단된다.

• Journal of Ginseng Research
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• 제40권1호
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• pp.28-37
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• 2016

Background: Panax ginseng cannot be cultivated on the same land consecutively for an extended period, and the underlying mechanism regarding microorganisms is still being explored. Methods: Polymerase chain reaction and denaturing gradient gel electrophoresis (PCR-DGGE) and BIO-LOG methods were used to evaluate the microbial genetic and functional diversity associated with the P. ginseng rhizosphere soil in various cultivation ages and modes. Results: The analysis of microbial diversity using PCR-DGGE showed that microbial communities were significantly variable in composition, of which six bacterial phyla and seven fungal classes were detected in P. ginseng soil. Among them, Proteobacteria and Hypocreales dominated. Fusarium oxysporum, a soilborne pathogen, was found in all P. ginseng soil samples except R0. The results from functional diversity suggested that the microbial metabolic diversity of fallow soil abandoned in 2003was the maximum and transplanted soil was higher than direct-seeding soil and the forest soil uncultivated P. ginseng, whereas the increase in cultivation ages in the same mode led to decreases in microbial diversity in P. ginseng soil. Carbohydrates, amino acids, and polymers were the main carbon sources utilized. Furthermore, the microbial diversity index and multivariate comparisons indicated that the augmentation of P. ginseng cultivation ages resulted in decreased bacterial diversity and increased fungal diversity, whereas microbial diversity was improved strikingly in transplanted soil and fallow soil abandoned for at least one decade. Conclusion: The key factors for discontinuous P. ginseng cultivation were the lack of balance in rhizosphere microbial communities and the outbreak of soilborne diseases caused by the accumulation of its root exudates.

• 생명과학회지
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• 제16권7호
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• pp.1112-1118
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• 2006

돼지 위축성 비염과 개의 kennel cough의 원인균인 B. bronchiseptica는 각 숙주의 상부 호흡기관의 점막에 집락을 형성하는 병원균으로서 철이 부족한 환경에서 hydroxamate type의 alcaligin이 라는 siderophore를 생산한다. Alcaligin의 생합성에 관련하는 구조유전자 중 alcA 유전자의 기능을 밝히고자 alcA 결손돌연변이주 구축을 통하여 확인하였다. alcA 유전자 결손 돌연변이를 위해 0.6 kb alcA 5' flanking DNA와 0.7 kb alcA 3' flanking DNA fragment들을 pCP1.11을 주형으로 하여 PCR법으로 증폭한 후, 5' flanking과 3' flanking DNA가 연결된 재조합 suicide vector pDMl을 구축하여 세포 접합을 통해 B. bronchiseptica로 도입시켰다. 도입된 pDM1으로부터 allelic exchange법에 의해 alcA 유전자가 결손된 돌연변이주 B. bronchiseptica H1을 얻을 수 있었다. B. bronchiseptica H1은 야생형인 B. bronchiseptica에 비하여 alcaligin siderophore를 거의 생성하지 못하였다. alc 오페론 중 promoter와 alcA 유전자만을 가지는 재조합 플라스미드를 B. bronchiseptica Hl에 도입하였을 때 alcaligin siderophore의 생산이 회복됨을 확인할 수 있었다. 이상의 결과로부터 alcA 유전자가 alcaligin 생합성에서 매우 중요한 역할을 수행하는 것을 알 수 있었다.

• Journal of Veterinary Science
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• 제22권5호
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• pp.63.1-63.14
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• 2021

Background: Recently, mesenchymal stem cells therapy has been performed in dogs, although the outcome is not always favorable. Objectives: To investigate the therapeutic efficacy of mesenchymal stem cells (MSCs) using dog leukocyte antigen (DLA) matching between the donor and recipient in vitro. Methods: Canine adipose-derived MSCs (cA-MSCs) isolated from the subcutaneous tissue of Dog 1 underwent characterization. For major DLA genotyping (DQA1, DQB1, and DRB1), peripheral blood mononuclear cells (PBMCs) from two dogs (Dogs 1 and 2) were analyzed by direct sequencing of polymerase chain reaction (PCR) products. The cA-MSCs were co-cultured at a 1:10 ratio with activated PBMCs (DLA matching or mismatching) for 3 days and analyzed for immunosuppressive (IDO, PTGS2, and PTGES), inflammatory (IL6 and IL10), and apoptotic genes (CASP8, BAX, TP53, and BCL2) by quantitative real-time reverse transcriptase-PCR. Results: cA-MSCs were expressed cell surface markers such as CD90⁺/44⁺/29⁺/45^- and differentiated into osteocytes, chondrocytes, and adipocytes in vitro. According to the Immuno Polymorphism Database, DLA genotyping comparisons of Dogs 1 and 2 revealed complete differences in genes DQA1, DQB1, and DRB1. In the co-culturing of cA-MSCs and PBMCs, DLA mismatch between the two cell types induced a significant increase in the expression of immunosuppressive (IDO/PTGS2) and apoptotic (CASP8/BAX) genes. Conclusions: The administration of cA-MSCs matching the recipient DLA type can alleviate the need to regulate excessive immunosuppressive responses associated with genes, such as IDO and PTGES. Furthermore, easy and reliable DLA genotyping technology is required because of the high degree of genetic polymorphisms of DQA1, DQB1, and DRB1 and the low readability of DLA 88.

• 한국육종학회지
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• 제40권4호
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• pp.387-393
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• 2008

Ds 삽입변이체로 부터 5,400개의 FST를 분석한 결과, intragenic FST가 48.1%로 2,597개, intergenic FST가 25.6%로 1,383개였으며 hot spot을 포함한 origin insertional sequence는 1,350개로 25%로 나타났다. Intragenic FST로서 선발된 2,597개를 이용하여 Ds와의 유전자형을 분석한 결과 53.6%인 1,393개가 heterozygous 혹은 homozygous 계통으로 나타났으며 이들에 대한 염색체상의 분포도는 3번 염색체에서 422 계통으로 가장 많은 분포도를 보여주었고 다른 염색체는 56 계통에서 157 계통 범위 내에 포함되었다. 1차적으로 유전자형 분석이 끝난 1,393개의 유전자들 중에서 expressed protein 등 알려지지 않은 것은 40.6%로 566개였으며 TIGR DB에서 염기서열의 유사성 검색을 통해 유전자의 명칭이 알려진 것은 59.4%인 827개로 나타났다.

• 한국해양생명과학회지
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• 제8권2호
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• pp.128-135
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• 2023

엽상자어(Leptocephalus)는 뱀장어목(Anguiliformes)에 속하는 어류의 자어로 5~6월에 우리나라 남해안 일대의 정치망에서 멸치와 함께 어획 후 방류하지만 대부분 폐사하는 실정이다. 따라서 본 연구의 목적은 멸치어업을 하는 정치망에서 무분별하게 포획되는 엽상자어의 종을 구명하여 수산자원보호 및 이들 자치어 생태 연구를 위한 기초 생물학적 자료를 제시하고자 수행하였다. 실험에 사용된 엽상자어는 5~6월에 남해의 정치망에서 채집하였으며, 외부형태와 유전학적 특성을 붕장어(Conger myriaster), 갯장어(Muraenesox cinereus) 및 뱀장어속(Anguilla) 4종의 성어와 비교하였다. 유전학적 분석은 추출한 DNA를 12s rRNA, 16s rRNA 부분단편을 PCR로 증폭하여 염기배열을 분석한 후 분자계통수를 작성하여 장어류 유생이 붕장어, 갯장어 및 뱀장어속 4종 중 어느 쪽의 성체와 클러스터 그룹화를 이루는지 계통학적 유연관계를 확인하였다. 엽상자어의 외부형태 계수 및 계측결과 엽상자어의 전장에 대한 머리길이의 백분비와 뒷지느러미 기점거리의 백분비는 붕장어와 가장 유사한 비율을 보였고, 척추골수 역시 붕장어와 가장 유사하였다. 또한 엽상자어의 유전자 분석결과는 모두 붕장어 성체와 클러스터 그룹화를 이루는 것을 확인함으로서 남해안에서 5~6월에 어획되는 엽상자어는 모두 붕장어의 자어임을 할 수 있었다.

• Journal of Ginseng Research
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• 제47권5호
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• pp.662-671
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• 2023

Background: 20(S)-protopanaxadiol (PPD), a ginsenoside metabolite, has prominent benefits for the central nervous system, especially in improving learning and memory. However, its transcriptional targets in brain tissue remain unknown. Methods: In this study, we first used mass spectrometry-based drug affinity responsive target stability (DARTS) to identify the potential proteins of ginsenosides and intersected them with the transcription factor library. Second, the transcription factor PURA was confirmed as a target of PPD by biolayer interferometry (BLI) and molecular docking. Next, the effect of PPD on the transcriptional levels of target genes of PURA in brain tissues was determined by qRT-PCR. Finally, bioinformatics analysis was used to analyze the potential biological features of these target proteins. Results: The results showed three overlapping transcription factors between the proteomics of DARTS and transcription factor library. BLI analysis further showed that PPD had a higher direct interaction with PURA than parent ginsenosides. Subsequently, BLI kinetic analysis, molecular docking, and mutations in key amino acids of PURA indicated that PPD specifically bound to PURA. The results of qRT-PCR showed that PPD could increase the transcription levels of PURA target genes in brain. Finally, bioinformatics analysis showed that these target proteins were involved in learning and memory function. Conclusion: The above-mentioned findings indicate that PURA is a transcription target of PPD in brain, and PPD upregulate the transcription levels of target genes related to cognitive dysfunction by binding PURA, which could provide a chemical and biological basis for the study of treating cognitive impairment by targeting PURA.

• Asian Pacific Journal of Cancer Prevention
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• 제13권12호
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• pp.6155-6157
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• 2012

Background: Mutations in the MAPK (Mitogen Activated Protein Kinase) signaling pathway - EGFR/Ras/RAF/MEK have been associated with the development of several carcinomas. ERK2, a downstream target of the MAPK pathway and a founding member of the MAPK family is activated by cellular signals emanating at the cell membrane. Activated ERK2 translocates into the nucleus to transactivate genes that promote cell proliferation. MKP - a dual specific phosphatase - interacts with activated ERK2 via the common docking (CD) domain of the later to inactivate (dephosphorylate) and effectively terminate further cell proliferation. A constitutively active form of ERK2 carrying a single point mutation - E322K in its CD domain, was earlier reported by our laboratory. In the present study, we investigated the prevalence of this CD domain E322K mutation in 88 well differentiated OSCC tissue samples. Materials and Method: Genomic DNA specimens isolated from 88 oral squamous cell carcinoma tissue samples were amplified with primers flanking the CD domain of the ERK2 gene. Subsequently, PCR amplicons were gel purified and subjected to direct sequencing to screen for mutations. Results: Direct sequencing of eighty eight OSCC samples identified an E322K CD domain mutation in only one (1.1%) OSCC sample. Conclusions: Our result indicates that mutation in the CD domain of ERK2 is rare in OSCC patients, which suggests the role of genetic alterations in other mitogenic genes in the development of carcinoma in the rest of the patients. Nevertheless, the finding is clinically significant, as the relatively rare prevalence of the E322K mutation in OSCC suggests that ERK2, being a common end point signal in the multi-hierarchical mitogen activated signaling pathway may be explored as a viable drug target in the treatment of OSCC.

• Asian Pacific Journal of Cancer Prevention
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• 제16권12호
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• pp.4915-4920
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• 2015

Background: : Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease mainly caused by mutations of the adenomatous polyposis coli (APC) gene with almost complete penetrance. These colorectal polyps are precancerous lesions that will inevitable develop into colorectal cancer at the median age of 40-year old if total proctocolectomy is not performed. So identification of APC germline mutations has great implications for genetic counseling and management of FAP patients. In this study, we screened APC germline mutations in Chinese FAP patients, in order to find novel mutations and the APC gene germline mutation characteristics of Chinese FAP patients. Materials and Methods: The FAP patients were diagnosed by clinical manifestations, family histories, endoscope and biopsy. Then patients peripheral blood samples were collected, afterwards, genomic DNA was extracted. The mutation analysis of the APC gene was conducted by direct polymerase chain reaction (PCR) sequencing for micromutations and multiplex ligation-dependent probe amplification (MLPA) for large duplications and/or deletions. Results: We found 6 micromutations out of 14 FAP pedigrees, while there were no large duplications and/or deletions found. These germline mutations are c.5432C>T(p. Ser1811Leu), two c.3926_3930delAAAAG (p.Glu1309AspfsX4), c.3921_3924delAAAA (p.Ile1307MetfsX13), c3184_3187delCAAA(p.Gln1061AspfsX59) and c4127_4126delAT (p.Tyr1376LysfsX9), respectively, and all deletion mutations resulted in a premature stop codon. At the same time, we found c.3921_3924delAAAA and two c.3926_3930delAAAAG are located in AAAAG short tandem repeats, c3184_3187delCAAA is located in the CAAA interrupted direct repeats, and c4127_4128 del AT is located in the 5'-CCTGAACA-3', 3'-ACAAGTCC-5 palindromes (inverted repeats) of the APC gene. Furthermore, deletion mutations are mostly located at condon 1309. Conclusions: Though there were no novel mutations found as the pathogenic gene of FAP in this study, we found nucleotide sequence containing short tandem repeats and palindromes (inverted repeats), especially the 5 bp base deletion at codon 1309, are mutations in high incidence area in APC gene,.

• Clinical and Experimental Reproductive Medicine
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• 제32권3호
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• pp.279-286
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• 2005

Objective: Although several genetic factors have been associated with defects in human spermatogenesis, the unambiguous causative genes have not been elucidated. The male infertility by haploinsufficiency of PRM1 or PRM2 has been reported in mouse model. The aim of this study was to identify the single nucleotide polymorphisms (SNPs) of PRM1 and PRM2, related to the genotype of Korean infertile men. Methods: Genomic DNAs were extracted from peripheral bloods of infertile men with oligozoospermia or azoospermia, and analyzed using polymerase chain reaction (PCR) and direct sequencing. We carried out the direct sequencing analysis of amplified fragments in PRM1 (557 nucleotides from -42 to 515) and PRM2 (599 nucleotides from 49 to 648) genes, respectively. Results: Three SNPs of coding region in the PRM1 gene was found in the analysis of 130 infertile men. However, the SNPs at a133g (aa 96.9%, ag 3.1% and gg 0.0%), c160a (cc 99.2%, ca 0.8% and aa 0.0%) and c321a (cc 56.9%, ca 35.4% and aa 7.7%) coded the same amino acids, in terms of silence phenotypes. On the other hand, as results of the PRM2 gene sequencing in 164 infertile men, only two SNPs, g398c (gg 62.2%, gc 31.1% and ga 6.7%) and a473c (aa 63.4%, ac 29.9% and cc 6.7%), were identified in the intron of the PRM2 gene. Conclusions: There was no mutation and significant SNPs on PRM1 and PRM2 gene in Korean infertile men. These results suggest that the PRM1 and PRM2 genes are highly conserved and essential for normal fertility of men.