• Title/Summary/Keyword: DNA 염기 서열

Search Result 1,579, Processing Time 0.032 seconds

Development of RAPD-SCAR Molecular Marker Related to Seed-hair Characteristic in Carrot (당근(Daucus carota var. sativa) 종자모 형질 관련 RAPD-SCAR 분자표지 개발)

  • Shim, Eun-Jo;Park, Sung-Kwan;Oh, Gyu-Dong;Jun, Sang-Jin;Park, Young-Doo
    • Horticultural Science & Technology
    • /
    • v.31 no.6
    • /
    • pp.756-763
    • /
    • 2013
  • Mechanical hair removal of carrot seed causes seed injuries and suppresses the germination in carrot cultivation. This study was performed to develop molecular markers for breeding high quality cultivars with short-hair seed. To meet this objective, random amplified polymorphic DNA (RAPD)-sequence characterized amplified region (SCAR) markers specifically linked to seed-hair characteristic were identified using CT-SMR 616 OP 389-1 line with short-haired seed and CT-SMR 616 OP 616-33 line with long-haired seed, bred by self-pollination for 6 years from 2008 to 2013, as parents. After seed hair lengths of these lines were analyzed using microscope, next generations were advanced and compared with the molecular markers polymorphism. From RAPD analysis using fixed lines in 2011, twelve RAPD primers showing polymorphic bands specific between the two lines were identified from 80 random primers. To develop RAPD-SACR marker, SCAR primers were designed based on sequence analysis of these specific RAPD bands and more than three combinations of primers were tested. As a result, it was found that the $SCA2_{1.2}$ amplified single polymorphic band from short-haired seed line. To confirm this result, $SCA2_{1.2}$ marker was retested by applying to the 2012 and 2013 progenies. Finally, it was concluded that the developed $SCA2_{1.2}$ marker distinguished short-haired line from long-haired seed line. Therefore, SCAR marker, $SCA2_{1.2}$ is expected to be utilized for breeding of the short-haired seed cultivars.

Physiological and Biochemical Characterization of Bacillus spp. from Polychaete, Perinereis aibuhitensis (갯지렁이(Perinereis aibuhitensis)에서 분리한 Bacillus spp.의 생리생화학적 특성 분석)

  • Shin, Seyeon;Yundendorj, Khorloo;Lee, Sang-Suk;Kang, Kyoung-Ho;Kahng, Hyung-Yeel
    • Journal of Life Science
    • /
    • v.23 no.3
    • /
    • pp.415-425
    • /
    • 2013
  • This study compared the characteristics of five Bacillus strains capable of aerobic and anaerobic growth, CBW3, CBW4, CBW9, CBW14 and EBW10. They were isolated and selected from a polychaete, Perinereis aibuhitensis, which is known as a good degrader of organic compounds in marine wetland. Based on a 16S rRNA sequence, CBW3 and CBW14 were found to share more than 99.8% similarity with B. nanhaiensis, B. arsenicus and B. barbaricus. CBW4, CBW9 and EBW10 shared 92.7%, 99.8%, and 99.8% similarity with B. anthracis, B. algicoa and B. thuringiensis, respectively. The temperature, salinity, and pH ranges of the cell growth of the Bacillus strains were $4-45^{\circ}C$, 0-17%, and pH 5-pH 9, respectively. All Bacillus strains were found to exhibit enzyme activities for the degradation of casein and starch. Notably, strain EBW10 exhibited the enzyme activities for all the tested macromolecules, DNA, casein, starch, cellulose, and four kinds of Tweens, which suggests the possibility that it had protease, amylase, cellulose, and lipase. All five Bacillus strains had alkaline phosphatase activities, and the strains CBW3, CBW4, and EBW10 also had acid phospatase. Strains CBW3 and EBW10 exhibited the enzyme activities both for esterase (C4) and esterase lipase (C8). The analysis of fatty acids revealed that in all strains, major fatty acids were anteiso $C_{15:0}$ and iso $C_{15:0}$.

Regulatory Characterization of xylA Promoter Region in Escherichia coli (대장균의 xylA 프로모터 영역의 조절 특성)

  • Kang, Byung-Tae;Roh, Dong-Hyun;Joo, Gil-Jae;Rhee, In-Koo
    • Applied Biological Chemistry
    • /
    • v.39 no.6
    • /
    • pp.443-448
    • /
    • 1996
  • In order to investigate the function of xylA promoter(Pxyl) as regulatory region Pxyl-lacZ fusion gene was constructed by the insertion of xylA promoter to the multiple cloning site of upstream of lacZ gene in a multicopy numbered plasmid pMC1403 containing promoterless lac operon, which was designated pMCX191, and Pxyl-lacZ fragment from pMCX191 was inserted to low copy numbered plasmid pLG339, designated pLGX191. The expressions of ${\beta}-galactosidase$ in these recombinant plasmids containing Pxyl-lacZ fusion gene were induced strongly by the addition of xylose, repressed by the addition of 0.2% glucose in the presence of xylose. The catabolite repressions were derepressed by the addition of 1 mM cAMP as same as native xylA gene. The fragment of xylA promoter was partially deleted from the upstream of xylA promoter by exonuclease III to investigate the regulation site of xylA promoter and the degrees of deletion derivatives of xylA promoter were analyzed by the DNA base sequencing. By the investigations of the induction by xylose, repression by glucose and derepression by cAMP on xylose isomerase production, the regulation site of xylA promoter may be located in segment between -165 and -59 bp upstream from the initiation site of xylA translation.

  • PDF

Species Identification of Noctuid Potential Pests of Soybean and Maize, and Estimation of Their Annual Adult Emergence in Suwon, Korea (수원지방에서 콩과 옥수수 가해 밤나방과의 잠재해충에 대한 종 동정과 연중 성충 발생 추정)

  • Jung, Jin Kyo;Kim, Eun Young;Kim, I Hyeon;Seo, Bo Yoon
    • Korean journal of applied entomology
    • /
    • v.59 no.2
    • /
    • pp.93-107
    • /
    • 2020
  • Adults of seven noctuid potential pests (Spodoptera frugiperda, S. litura, S. exigua, Ctenoplusia agnata, Mythimna loreyi, Athetis dissimilis, and A. lepigone) of soybean and maize in Suwon, Korea were identified by their morphological characteristics in the wing pattern and male genitalia and partial mitochondrial DNA sequences of cytochrome c oxidase subunit 1 gene. The generation number of adults that emerge annually in six species (except A. lepigone) was estimated from the data on density fluctuations of adults caught in sex pheromone traps in 2019 and the forecasted data using temperature-associated development and reproduction models for those species. S. frugiperda adults were caught from July 27th to October 31st in 2019, and hence were initially estimated to emerge three times per year. But, it was finally expected that S. frugiperda adults could possibly emerge a total of four times per year in Suwon, considering larval emergence observed during mid- and late June in other areas. Adult emergence of S. litura, S. exigua, C. agnata, and M. loreyi in 2019 was observed from May 29th to November 6th, from May 14th to November 6th, from May 26th to October 25th, and from May 31st to November 23rd, respectively. Annual adult emergence of these four species was estimated as at least four times. Adults of A. dissimilis were caught from May 26th to September 11th in 2019, and adult emergence was estimated at only twice per annum. It was postulated that the first adult populations of five species except the two Athetis species were probably migrated from other areas.

Process Optimization of Ginseng Berry Extract Fermentation by Lactobacillus sp. Strain KYH isolated from Fermented Kimchi and Product Analysis (발효 김치로부터 분리한 Lactobacillus sp. Strain KYH를 이용한 진생베리 추출물 최적 발효 공정 확립 및 생성물의 특성 분석)

  • Ha, Yoo-Jin;Yoo, Sun-Kyun;Kim, Mee Ree
    • Journal of the East Asian Society of Dietary Life
    • /
    • v.26 no.1
    • /
    • pp.88-98
    • /
    • 2016
  • The pharmacological effects of ginseng berry have been known to improve psychological function, immune activities, cardiovascular conditions, and certain cancers. It is also known that fermentation improves the bioavailability of human beneficial natural materials. Accordingly, we investigated the optimal fermentation conditions of ginseng berry extract with strain isolated from conventional foods. We also analyzed the fermentation product and its antioxidant activity. The bacterium isolated from fermented kimchi was identified as Lactobacillus sp. strain KYH. To optimize the process, fermentation was performed in a 5 L fermenter containing 3 L of ginseng berry extract at 200 rpm for 72 hr. Under optimized conditions, batch and fed-batch fermentations were performed. After fermentation, organic acids, amino acids, sugars, ginsenosides, and antioxidant activity were evaluated. The optimum fermentation conditions were determined as pH 7.0 and a temperature of $30^{\circ}C$, respectively. After fermentation, the amounts and compositions of organic acids, amino acids, sugars, ginsenosides, and antioxidant activity were altered. In comparing the distribution of ginsenosides with that before fermentation, the ginsenoside Re was a major product. However, amounts of ginsenosides Rb1, Rc, and Rd were reduced, whereas amounts of ginsenosides Rh1 and Rh2 increased. Total phenol content increased to 43.8%, whereas flavonoid content decreased to 19.8%. The DPPH radical scavenging activity and total antioxidant activity increased to 27.2 and 19.4%, respectively.

Enhanced production of monacolin-K through supplement of monacolin-K precursors into production medium and cloning of SAM synthetase gene (metK) (Precursor제공 및 생합성 관련 유전자의 cloning을 통한 Monacolin-K 생산성 향상)

  • Lee, Mi-Jin;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
    • /
    • v.23 no.6
    • /
    • pp.519-524
    • /
    • 2008
  • Monacolin-K is a strong anti-hypercholesterolemic agent produced by Monascus sp. via polyketide pathway. High-yielding mutants of monacolin-K were developed through rational screening strategies adopted based on understanding of monacolin-K biosynthetic pathway. Through the experiments for investigating various amino acids as putative precursors for the monacolin-K biosynthesis, it was found that production level of monacolin-K was remarkably increased when optimum amount of cysteine was supplemented into the production medium. We suggested that these phenomena might be related to the special roles of SAM (S-adenosyl methionine), a putative methyl group donor in the biosynthetic pathway of monacolin-K, demonstrating close interrelationship between SAM-synthesizing primary metabolism and monacolin-K synthesizing secondary metabolism. Namely, increase in the intracellular amount of SAM derived from the putative precursor, cysteine which was extracellularly supplemented into the production medium might contribute to the significant enhancement in the monacolin-K biosynthetic capability of the highly mutated producers. On the basis of these assumptions derived from the above fermentation results, we decided to construct efficient expression vectors harboring SAM synthetase gene (metK) cloned from A. nidulans, with the hope that increased intracellular level of SAM could lead to further enhancement in the monacolin-K production through overcoming a rate-limiting step associated with monacolin-K biosynthesis. Hence, in order to overcome the plausible rate-limiting step associated with monacolin-K biosynthesis by increasing intracellular level of SAM, we transformed the producer mutants with an efficient expression vector harboring gpdA promoter of the producer microorganism, and metK gene. Notably, from the resulting various transformants, we were able to screen a very high-yielding transformant which showed approximately 3.3 fold higher monacolin-K productivity than the parallel nontransformed mutants in shake flask cultures performed under the identical fermentation conditions.

Development of Detection Method for Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus using 16S rRNA Gene (16S rRNA를 이용한 다금바리, 자바리, 능성어 판별법 개발)

  • Park, Yong-Chjun;Jung, Yong-Hyun;Kim, Mi-Ra;Shin, Joon-Ho;Kim, Kyu-Heon;Lee, Jae-Hwang;Cho, Tae-Yong;Lee, Hwa-Jung;Lee, Sang-Jae;Han, Sang-Bae
    • Korean Journal of Food Science and Technology
    • /
    • v.45 no.1
    • /
    • pp.1-7
    • /
    • 2013
  • Niphon spinosus, Epinephelus bruneus, and Epinephelus septemfasciatus are involved in the Perciformes Order and Serranidae Family. When E. bruneus and E. septemfasciatus are fully grown, the striped pattern on the body gradually disappears. Therefore, morphological classification of adult fishes is quite difficult to identify the differences to N. spinosus. In this study, we investigate the method to differentiate those using PCR. To design the primers, 16S rRNA region of N. spinosus, E. bruneus, and E. septemfasciatus registered in the GeneBank (www.ncbi.nlm.nih.gov) have been used and for the analysis, Bio Edit ver. 7.0.9.0 was used. As a result, it was design NS-003-F/NS-005-R (136 bp), EB-001-F/EB-002-R (181 bp), and ES-001-F/ES-001-R (123 bp) primers for the differentiation of each 3 different fishes. Therefore, the species-specific primer sets would be a useful tool for scientific and speedy differentiation against the illegal distribution for consumer protection.

Development of Method using LC-ESI-MS/MS and KASP for Identification of Gymnema sylvestre in Food (식품에서 당살초 판별을 위한 LC-ESI-MS/MS 분석법과 KASP 마커 개발)

  • Park, Boreum;Lee, Sun Hee;Eom, Kwonyong;Noh, Eunyoung;Moon Han, Kyoung;Hwang, Jinwoo;Kim, Hyungil;Baek, Sun Young
    • Journal of Food Hygiene and Safety
    • /
    • v.37 no.2
    • /
    • pp.46-54
    • /
    • 2022
  • Known for its effectiveness in weight loss and diabetes prevention, Gymnema sylvestre products can be found in the US, Japanese, and Indian markets. However, the recommended dosage or safety of these products has not yet been proven. Therefore, development of an analytical method for detecting the content of Gymnema sylvestre in food products is required. Accordingly, this study proposes an analysis method that can examine Gymnema sylvestre in food using LC-ESI-MS/MS and KASP (Kompetitive Allele-Specific PCR) markers. In LC-ESI-MS/MS, a simultaneous analysis method for gymnemic acid and deacylgymnemic acid was optimized using negative ionization mode, and its validation test was completed for solid and liquid samples. In addition, KASP markers were prepared by finding the specific SNP of G. sylvestre in ITS2 and matK through DNA barcodes. The two KASP markers returned positive FAM fluorescence result when combined with G. sylvestre, and this aspect was confirmed in raw G. sylvestre as well. The applicability of the method was tested on 21 different food and healthy functional products containing G. sylvestre purchased on the internet. As a result, although there was a difference in the ratios of gymnemic acid and deacylgymnemic acid in LC-ESI-MS/MS, the index component was detected in all 21 products samples. In the KASP analysis, 9 products returned positive FAM result, and the rest of the products were found to be containing G. sylvestre extract. This study is the first study to use the dual system of LC-ESI-MS/MS and KASP for the analysis of G. sylvestre. The study has confirmed that these two methods are applicable to the examine G. sylvestre content in food products.

Review of the Korean Indigenous Species Investigation Project (2006-2020) by the National Institute of Biological Resources under the Ministry of Environment, Republic of Korea (한반도 자생생물 조사·발굴 연구사업 고찰(2006~2020))

  • Bae, Yeon Jae;Cho, Kijong;Min, Gi-Sik;Kim, Byung-Jik;Hyun, Jin-Oh;Lee, Jin Hwan;Lee, Hyang Burm;Yoon, Jung-Hoon;Hwang, Jeong Mi;Yum, Jin Hwa
    • Korean Journal of Environmental Biology
    • /
    • v.39 no.1
    • /
    • pp.119-135
    • /
    • 2021
  • Korea has stepped up efforts to investigate and catalog its flora and fauna to conserve the biodiversity of the Korean Peninsula and secure biological resources since the ratification of the Convention on Biological Diversity (CBD) in 1992 and the Nagoya Protocol on Access to Genetic Resources and the Fair and Equitable Sharing of Benefits (ABS) in 2010. Thus, after its establishment in 2007, the National Institute of Biological Resources (NIBR) of the Ministry of Environment of Korea initiated a project called the Korean Indigenous Species Investigation Project to investigate indigenous species on the Korean Peninsula. For 15 years since its beginning in 2006, this project has been carried out in five phases, Phase 1 from 2006-2008, Phase 2 from 2009-2011, Phase 3 from 2012-2014, Phase 4 from 2015-2017, and Phase 5 from 2018-2020. Before this project, in 2006, the number of indigenous species surveyed was 29,916. The figure was cumulatively aggregated at the end of each phase as 33,253 species for Phase 1 (2008), 38,011 species for Phase 2 (2011), 42,756 species for Phase 3 (2014), 49,027 species for Phase 4 (2017), and 54,428 species for Phase 5(2020). The number of indigenous species surveyed grew rapidly, showing an approximately 1.8-fold increase as the project progressed. These statistics showed an annual average of 2,320 newly recorded species during the project period. Among the recorded species, a total of 5,242 new species were reported in scientific publications, a great scientific achievement. During this project period, newly recorded species on the Korean Peninsula were identified using the recent taxonomic classifications as follows: 4,440 insect species (including 988 new species), 4,333 invertebrate species except for insects (including 1,492 new species), 98 vertebrate species (fish) (including nine new species), 309 plant species (including 176 vascular plant species, 133 bryophyte species, and 39 new species), 1,916 algae species (including 178 new species), 1,716 fungi and lichen species(including 309 new species), and 4,812 prokaryotic species (including 2,226 new species). The number of collected biological specimens in each phase was aggregated as follows: 247,226 for Phase 1 (2008), 207,827 for Phase 2 (2011), 287,133 for Phase 3 (2014), 244,920 for Phase 4(2017), and 144,333 for Phase 5(2020). A total of 1,131,439 specimens were obtained with an annual average of 75,429. More specifically, 281,054 insect specimens, 194,667 invertebrate specimens (except for insects), 40,100 fish specimens, 378,251 plant specimens, 140,490 algae specimens, 61,695 fungi specimens, and 35,182 prokaryotic specimens were collected. The cumulative number of researchers, which were nearly all professional taxonomists and graduate students majoring in taxonomy across the country, involved in this project was around 5,000, with an annual average of 395. The number of researchers/assistant researchers or mainly graduate students participating in Phase 1 was 597/268; 522/191 in Phase 2; 939/292 in Phase 3; 575/852 in Phase 4; and 601/1,097 in Phase 5. During this project period, 3,488 papers were published in major scientific journals. Of these, 2,320 papers were published in domestic journals and 1,168 papers were published in Science Citation Index(SCI) journals. During the project period, a total of 83.3 billion won (annual average of 5.5 billion won) or approximately US $75 million (annual average of US $5 million) was invested in investigating indigenous species and collecting specimens. This project was a large-scale research study led by the Korean government. It is considered to be a successful example of Korea's compressed development as it attracted almost all of the taxonomists in Korea and made remarkable achievements with a massive budget in a short time. The results from this project led to the National List of Species of Korea, where all species were organized by taxonomic classification. Information regarding the National List of Species of Korea is available to experts, students, and the general public (https://species.nibr.go.kr/index.do). The information, including descriptions, DNA sequences, habitats, distributions, ecological aspects, images, and multimedia, has been digitized, making contributions to scientific advancement in research fields such as phylogenetics and evolution. The species information also serves as a basis for projects aimed at species distribution and biological monitoring such as climate-sensitive biological indicator species. Moreover, the species information helps bio-industries search for useful biological resources. The most meaningful achievement of this project can be in providing support for nurturing young taxonomists like graduate students. This project has continued for the past 15 years and is still ongoing. Efforts to address issues, including species misidentification and invalid synonyms, still have to be made to enhance taxonomic research. Research needs to be conducted to investigate another 50,000 species out of the estimated 100,000 indigenous species on the Korean Peninsula.