• Title/Summary/Keyword: MtDNA

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Pepper Blight Disease Inhibition Metagenome Clone Screening Using Soil Metagenome Library (토양 Metagenome Library로부터 고추역병 저해 클론 탐색)

  • Park, Hae-Chul;Sung, So-Ra;Kim, Dong-Gwan;Koo, Bon-Sung;Jeong, Byeong-Moon;Kim, Jin-Heung;Yoon, Moon-Young
    • Korean Journal of Microbiology
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    • v.45 no.2
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    • pp.228-231
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    • 2009
  • We have purified Phytophthora capsici alpha and beta tubulin from Escherchia coli BL21(DE3). The recombinant alpha and beta tubulins were assembled into microtubule in vitro with specific conditions. The metagenome library was isolated from soil in the Mt. Yeo-Ki, Suwon, Korea and manufactured with the method mentioned in experiment contents for in vitro screening of microtubule assembly screening. FRET effect was used for microtubule assembly inhibitor screening with metagenome library. We got 2 metagenome clones from in vitro screening, and these 2 hit clones showed P. capsici growth inhibition activity on the growing pepper plants. These results suggest that new development of potent inhibitor for pepper blight disease and new approach to prevention of pepper blight disease.

Possibility of Natural Hybridization between Red Seabream (Pagrus major) and Blackhead Seabream (Acanthopagrus schlegeli) (참돔(Pagrus major)과 감성돔(Acanthopagrus schlegeli) 종간 자연 잡종 가능성)

  • Kang, Jung-Ha;Yang, Sang-Geun;Kim, Eun-Mi;Noh, Eun-Soo;Kim, Dong-Gyun;Kim, Bong-Seok;Choi, Tae-Jin
    • Journal of Life Science
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    • v.25 no.1
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    • pp.16-20
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    • 2015
  • During the storage of these two species in a large conservation tank, fertilized eggs were collected and the offspring were raised. During culturing of the offspring, individuals with mixed characteristics of these two species were observed, and 96 individuals were randomly tested using microsatellite markers applicable to both species. Among the 96 individuals, 15 individuals with mixed morphological characteristics were confirmed to be hybrids showing both of genotypes red seabream and blackhead seabream. Additionally, based on sequence analysis of mitochondrial cytochrome oxidase subunit 1 (mtDNA CO1), 81 showed 99% nucleotide sequence identity to that of black sea bream, and the remaining 15 individuals showed over 99% sequence identity to that of red seabream. So, hybrids were produced by female red seabream and male blackhead seabream. These results suggest that hybrids may form in nature between these two species if their habitats overlap due to the influence of humans or global climate change.

The Detection and a Quantitative Evaluation of Viable but Non-Culturable Soil Bacteria Using a Modified Direct Viable Count Method (변형된 DVC법을 이용한 난배양성 토양세균의 검출 및 정량적 평가)

  • 황경숙;양희찬;염곡효
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.181-186
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    • 2003
  • This study was performed to analyze quantitatively the number of living bacteria in forest soil samples collected from Mt. Keryong using improved direct viable count (DVC) and plate count (PC) methods. The number of living bacteria by DVC comprised 18~44% of the total direct count (TDC), whereas the number of living bacteria by PC was less than 1% of TDC. These results showed that viable but non-culturable (VBNC) bacteria existed in the soil with high percentages. Besides, DVC was proved to make it possible to make a quantitative detection of the VBNC bacteria. On the other hand, upon measuring the value from the conventional nutrient broth (NB) and $10^{-2}$ folded diluted nutrient broth (DNB), the values from the DNB showed 5 to 10 times higher than those from the conventional NB medium. These results indicate that oligotrophic bacterial groups, which could multiply in the low nutrient broth, abundantly exist in the soil ecosystem. It would also be possible to apply this kind of method to other substrate to make a quantitative detection of soil bacterial groups.

Ontogenetic Color Variation of Abudefduf notatus (Pomacentridae: Perciformes) Revealed by 16S rRNA Sequences Analysis (미토콘드리아 16S rRNA 염기서열 분석에 의해 밝혀진 동갈자돔 치어의 성장에 따른 체색변이)

  • Song, Young Sun;Kim, Jin-Koo
    • Korean Journal of Ichthyology
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    • v.25 no.2
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    • pp.53-58
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    • 2013
  • Seven individuals (16.1~29.1 mm in length) which were estimated with fishes of genus Abudefduf were collected in Seogwipo, Jeju Island in the summer of 2011 and 2012. Among them, five individuals (20.8~29.1 mm SL) are similar to Abudefduf notatus, based on morphological characters such as yellow transverse band on the body, one small black spot on the base of the pectoral fin, and yellow caudal fin. On the other hand, two individuals (16.1 mm, 17.0 mm SL) differ from them in several light transverse bands and transparent rays of all fins except for pelvic fin, and anterior transparent head with only one (16.1 mm). According to the results of molecular analyses of 578 base pairs of mtDNA 16S rRNA sequences, these individuals correspond to A. notatus adult, with a high bootstrap value of 95% (genetic distance, d=0.002). Therefore, this study shows that the individuals more than 20.8 mm similar to adult body color but the 16.1 mm individual differs to that of adult. We confirm that this species changes to body color during their early life stages. This result regards as a survival strategy to protect themselves against their predator during their early life stages.

The Mitochondrial Warburg Effect: A Cancer Enigma

  • Kim, Hans H.;Joo, Hyun;Kim, Tae-Ho;Kim, Eui-Yong;Park, Seok-Ju;Park, Ji-Kyoung;Kim, Han-Jip
    • Interdisciplinary Bio Central
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    • v.1 no.2
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    • pp.7.1-7.7
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    • 2009
  • "To be, or not to be?" This question is not only Hamlet's agony but also the dilemma of mitochondria in a cancer cell. Cancer cells have a high glycolysis rate even in the presence of oxygen. This feature of cancer cells is known as the Warburg effect, named for the first scientist to observe it, Otto Warburg, who assumed that because of mitochondrial malfunction, cancer cells had to depend on anaerobic glycolysis to generate ATP. It was demonstrated, however, that cancer cells with intact mitochondria also showed evidence of the Warburg effect. Thus, an alternative explanation was proposed: the Warburg effect helps cancer cells harness additional ATP to meet the high energy demand required for their extraordinary growth while providing a basic building block of metabolites for their proliferation. A third view suggests that the Warburg effect is a defense mechanism, protecting cancer cells from the higher than usual oxidative environment in which they survive. Interestingly, the latter view does not conflict with the high-energy production view, as increased glucose metabolism enables cancer cells to produce larger amounts of both antioxidants to fight oxidative stress and ATP and metabolites for growth. The combination of these two different hypotheses may explain the Warburg effect, but critical questions at the mechanistic level remain to be explored. Cancer shows complex and multi-faceted behaviors. Previously, there has been no overall plan or systematic approach to integrate and interpret the complex signaling in cancer cells. A new paradigm of collaboration and a well-designed systemic approach will supply answers to fill the gaps in current cancer knowledge and will accelerate the discovery of the connections behind the Warburg mystery. An integrated understanding of cancer complexity and tumorigenesis is necessary to expand the frontiers of cancer cell biology.

Sequence Diversity of Mitochondrial Cytochrome b Gene in Grey Goral Naemorhedus caudatus(Artiodactyla, Bovidae) from Korea (한국의 산양(우제목, 소과)의 미토콘드리아 Cytochrome b 염기서열 다양성)

  • Koh, Hung-Sun;Yang, Byong-Guk;Lee, Bae-Kun;Lee, Jong-Hyong
    • Animal Systematics, Evolution and Diversity
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    • v.18 no.1
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    • pp.13-21
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    • 2002
  • To add genetic information to the conservation efforts on grey coral (Naemorhedus caudatus) in Korea, we investigated the pattern of mitochondrial cytochrome b gene sequence (606 bp) of six specimens from two localities in Korea. The corresponding sequences of N. caudatus in China obtained from GenBank were also used. The nucleotide Tamura-Nei distances between each of four haplotypes of N. caudatus in Korea and the haplotype of N. caudatus in China varied from 0.0650 to 0.0803: N. caudatus revealed high level of sequence diversity in Bovidae. In N. caudatus in Korea, the distances among three haplotypes at Yanggu were 0.0151 to 0.0185, and it suggests that the genetic diversity of Yanggu population was decreased in low level. Moreover, the distances between each of three haplotypes at Yanggu and one haplotype at Samcheok were 0.0343 to 0.0489. It indicates that habitat isolation caused the continuous increase of genetic distance with geographic distance in N. caudatus, and various conservation plans for mitigating the loss of genetic diversity in Korea have to be in immediate action. To clarify the taxonomic status of N. caudatus, the sequence (276 bp) of N. goral available from GenBank were also utilized, and n goral was not distinct from N. caudatus. It suggests that they may be conspecific, but further analyses with additional specimens of two species are necessary.

Identification of Pleurotus ostreatus cultivars with the application of multiplex-simple sequence repeat markers (Multiplex SSR마커를 이용한 느타리(Pleurotus ostreatus) 품종 판별)

  • Choi, Jong In;Jung, Hwa Jin;Na, Kyeong sook;Oh, Min-Ji;Kim, Min-Keun;Ryu, Jae-San
    • Journal of Mushroom
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    • v.19 no.1
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    • pp.76-80
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    • 2021
  • To develop a method for the differentiation of Pleurotus ostratus cultivars, the multiplex-simple sequence repeat (SSR) primer set based on the SSRs obtained from whole genomic DNA sequence analysis was designed with two polymerase chain reaction (PCR) primer sets. These SSR primer sets were employed to distinguish 10 cultivars and strains. Twenty polymorphic markers were selected based on the genotyping results. PCR with each primer produced 1-4 distinct bands ranging in size from 150 to 350 bp, which was within the expected range. However, since a sole SSR marker was unable to detect polymorphisms in every cultivar, multiplex PCRs with composite PCR primer sets were employed. The multiplex primer, "166+115," completely discriminated 12 cultivars and strains with 40 loci, which were 12 more than the simple arithmetic addition of each locus of the primers 115 and 166. These results might be useful to provide an efficient method for the differentiation of P. ostreatus cultivars with separate PCRs for the quality control of spawn and protection of breeders' rights.

Clinicopathologic Characteristics and Prognoses for Multicentric Occurrence and Intrahepatic Metastasis in Synchronous Multinodular Hepatocellular Carcinoma Patients

  • Li, Shi-Lai;Su, Ming;Peng, Tao;Xiao, Kai-Yin;Shang, Li-Ming;Xu, Bang-Hao;Su, Zhi-Xiong;Ye, Xin-Ping;Peng, Ning;Qin, Quan-Lin;Chen, De-Feng;Chen, Jie;Li, Le-Qun
    • Asian Pacific Journal of Cancer Prevention
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    • v.14 no.1
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    • pp.217-223
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    • 2013
  • Background: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and the outcomes for patients are still poor. It is important to determine the original type of synchronous multinodular HCC for preoperative assessment and the choice of treatment therapy as well as for the prediction of prognosis after treatment. Aims: To analyze clinicopathologic characteristics and prognoses in patients with multicentric occurrence (MO) and intrahepatic metastasis (IM) of synchronous multinodular hepatocellular carcinoma (HCC). Methods: The study group comprised 42 multinodular HCC patients with a total of 112 nodules. The control group comprised 20 HCC patients with 16 single nodular HCC cases and 4 HCC cases with a portal vein tumor emboli. The mitochondrial DNA (mtDNA) D-loop region was sequenced, and the patients of the study group were categorized as MO or IM based on the sequence variations. Univariate and multivariate analyses were used to determine the important clinicopathologic characteristics in the two groups. Results: In the study group, 20 cases were categorized as MO, and 22 as IM, whereas all 20 cases in the control group were characterized as IM. Several factors significantly differed between the IM and MO patients, including hepatitis B e antigen (HBeAg), cumulative tumor size, tumor nodule location, cirrhosis, portal vein and/or microvascular tumor embolus and the histological grade of the primary nodule. Multivariate analysis further demonstrated that cirrhosis and portal vein and/or microvascular tumor thrombus were independent factors differentiating between IM and MO patients. The tumor-free survival time of the MO subjects was significantly longer than that of the IM subjects ($25.7{\pm}4.8$ months vs. $8.9{\pm}3.1$ months, p=0.017). Similarly, the overall survival time of the MO subjects was longer ($31.6{\pm}5.3$ months vs. $15.4{\pm}3.4$ months, p=0.024). The multivariate analysis further demonstrated that the original type (p=0.035) and Child-Pugh grade (p<0.001) were independent predictors of tumor-free survival time. Cirrhosis (p=0.011), original type (p=0.034) and Child-Pugh grade (p<0.001) were independent predictors of overall survival time. Conclusions: HBeAg, cumulative tumor size, tumor nodule location, cirrhosis, portal vein and/or microvascular tumor embolus and histological grade of the primary nodule are important factors for differentiating IM and MO. MO HCC patients might have a favorable outcome compared with IM patients.

Detection and Differentiation of Intentional and Unintentional Mixture in Raw Meats Using Real-time PCR (Real-time PCR을 이용한 식육원료의 의도적, 비의도적 혼입 판별법 개발)

  • Kim, Kyu-Heon;Kim, Mi-Ra;Park, Young-Eun;Kim, Yong-Sang;Lee, Ho-Yeon;Park, Yong-Chjun;Kim, Sang Yub;Choi, Jang Duck;Jang, Young-Mi
    • Journal of Food Hygiene and Safety
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    • v.29 no.4
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    • pp.340-346
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    • 2014
  • In this study, the detection method was developed using real-time PCR to distinguish 4 species (bovine, porcine, horse, and chicken) of raw meats. The genes for distinction of species about meats targeted at 12S rRNA and 16S rRNA parts in mitochondrial DNA. Probes were designed to have a 5' FAM and a TAMRA at the 3' end. This study is to develop 4 species-specific primer and probes about raw materials and real-time PCR on 10 strains to observe the products of non-specific signal for similar species. As a result, any non-specific signal were not detected among each other. Real-time PCR method was developed for quantitation and identification of intentional and unintentional mixture in ground mixed meat (The difference of $C_T$ value between intentional mixture and 100% meat: $${\leq_-}$$ cycles, The difference of $C_T$ value between unintentional mixture and 100% meat: $${\geq_-}$$ cycles). The detection and differentiation of intentional and unintentional mixture in this study would be applied to food safety management for eradication of adulterated food distribution and protection of consumer's right.

Analysis of Genetic Characteristics and Probability of Individual Discrimination in Korean Indigenous Chicken Brands by Microsatellite Marker (MS 마커를 이용한 토종닭 브랜드의 유전적 특성 및 개체 식별력 분석)

  • Suh, Sangwon;Cho, Chang-Yeon;Kim, Jae-Hwan;Choi, Seong-Bok;Kim, Young-Sin;Kim, Hyun;Seong, Hwan-Hoo;Lim, Hyun-Tae;Cho, Jae-Hyeon;Ko, Yeoung-Gyu
    • Journal of Animal Science and Technology
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    • v.55 no.3
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    • pp.185-194
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    • 2013
  • Microsatellite markers have been a useful genetic tool in determining diversity, relationships and individual discrimination studies of livestock. The level of genetic diversity, relationships among two Korean indigenous chicken brand populations (Woorimatdag: WR, Hanhyup3: HH) as well as two pure populations (White Leghorn: WL, Rhode Island Red: RIR) were analyzed, based on 26 MS markers. A total of 191 distinct alleles were observed across the four chicken populations, and 47 (24.6%) of these alleles were unique to only one population. The mean $H_{Exp}$ and PIC were estimated as 0.667 and 0.630. Nei's $D_A$ genetic distance and factorial correspondence analysis (FCA) showed that the four populations represented four distinct groups. However, the genetic distance between each Korean indigenous chicken brand (WR, HH) and the pure population (WL, RIR) were threefold that among the WR and HH. For the STRUCTURE analyses, the most appropriate number of clusters for modeling the data was determined to be three. The expected probabilities of identity among genotypes of random individuals (PI) were calculated as $1.17{\times}10^{-49}$ (All 26 markers) and $1.14{\times}10^{-15}$, $7.33{\times}10^{-20}$ (9, 12 with the highest PI value, respectively). The results indicated that the brand chicken breed traceability system employing the own highest PI value 9 to 12 markers, and might be applicable to individual identification of Korean indigenous chicken brand.