• Title/Summary/Keyword: DNA screening

Search Result 708, Processing Time 0.028 seconds

Screening and Characteristics of Useful Fungi for Brewing from Commercial Nuruk in Chungcheong Provinces (충청지역 누룩에서 양조용 우수 곰팡이의 탐색 및 특성)

  • Baek, Seong-Yeol;Yun, Hye-Ju;Choi, Hye-Sun;Hong, Seung-Beom;Koo, Bon-Sung;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
    • /
    • v.38 no.4
    • /
    • pp.373-378
    • /
    • 2010
  • Studies on standardization and quality upgrade of nuruk which is a basic component in brewing are required to increase the quality level of Korean traditional rice wines and to develop the technology for practical use of it. It is important to isolate best strains, to improve the properties and effectively preserve them for brewing industry. In this study, 16 commercial nuruk samples were obtained from the commercial markets located in Chungcheong areas in Korea. 174 fungal strains were isolated from the samples on DG18 medium using a dilution plating method and then screened for enzyme activity and acid production. The active strains were identified based on the morphological characteristics and ITS sequence analysis. Out of 174 strains, 12 strains showed high amylase activity. Especially, Rhizopus sp. CN084, CN174, Aspergillus sp. CN161 and Mycocladus sp. CN042 showed high saccharogenic power and dextrinogenic enzyme activity on cooked wheat bran medium. On the other hand, Aspergillus sp. CN010, CN161, Rhizopus sp. CN105, CN168 and Rhizomucor sp. CN088 produced high acid production on the same medium. Our results showed that the active strains may be used as microbial sources for nuruk starter with good quality in brewing.

Prenatal Diagnosis of a Satellited Chromosome 8p Results from a de novo Cryptic Translocation between Chromosomes 8 and 22 (산전 진단에서 관찰된 8번과 22번 염색체 사이의 미세 전좌에 의한 8번 염색체 단완 위성체)

  • Oh, Ah-Rum;Lee, Bom-Yee;Choi, Ene-Yuong;Ryu, Hyun-Mee;Lee, Seung-Jae;Jung, Ji-Ye;Park, So-Yeon
    • Journal of Genetic Medicine
    • /
    • v.8 no.2
    • /
    • pp.135-138
    • /
    • 2011
  • The authors of the present study report the prenatal detection of a chromosomal abnormality with additional satellites on the distal short arm of chromosome 8. A 35-year-old woman was referred for amniocentesis because of her advanced maternal age and positive result for maternal serum screening test. Cytogenetic analysis of cultured amniocytes showed a satellite 8p chromosome. The satellite 8p chromosome was positive for nucleolus organizer region (NOR) staining. The parents' karyotypes were normal. Fluorescence in situ hybridization (FISH) study for metaphases of fetal amniocytes revealed a cryptic translocation of chromosomes 8p and 22p. The fetal karyotype was described as 46,XY,8ps.ish t(8;22)(p23.3;p11.2) (D8S504-;D8S504+)dn. The parents decided to continue the pregnancy and a phenotypically normal boy was born at 38 weeks of gestation. In case of de novo terminal NORs detected prenatally, more accurate cytogenetic and molecular analysis should be performed in order to rule out cryptic chromosomal rearrangement among other chromosomes.

Characterization of Cellulase from Bacillus subtilis NSC Isolated from Soil (토양으로부터 단리한 Bacillus subtilis NSC 유래 Cellulase의 특성 규명)

  • Kim, Sang Jin;Park, Chang-Su
    • Journal of Chitin and Chitosan
    • /
    • v.23 no.4
    • /
    • pp.228-233
    • /
    • 2018
  • We isolated microorganisms from soil, which is sampled at forest, Kyeonbuk, Korea, as cellulolytic microorganisms. The isolated strains were identified by analysis of 16S rRNA gene from the starins. The result, four kinds of Bacillus subtilis, one kind of Bacillus amyloliquefaciens, and one kind of Bacillus cereus were identified. Among these strains, Bacillus subtilis was selected due to its high cellulase activity and this strain was named as Bacillus subtilis CNS. The optimum pH and temperature of the cellulase from Bacillus subtilis CNS was pH 5.0 and $40^{\circ}C$, respectively. In the investigation of pH and temperature stability, the cellulase from Bacillus subtilis NSC stabled pH 4.0~6.0 range and until $40^{\circ}C$ for 30 min perfectly. In the enzyme activity for various cellulosic substrate, cellulase from Bacillus subtilis CNS showed the highest activity for CM-cellulose. And, the enzyme activities for alkali swollen cellulose, Alpha-cellulose, Sigmacell-cellulose, and Avicel were approximately 31%, 8%, 8% and 4% of activity for CM-cellulose, respectively. In the degradation of CM-cellulose, the 0.26 U/ml and 0.52 U/ml of cellulase showed 0.43 and 0.76 U/ml activity for CM-cellulose after the reaction of 120 min, respectively.

Establishment of Plasma Working Standards for the Performance and Quality Assurance of NAT Screening Tests for HIV, HCV and HBV (HIV, HCV와 HBV 유전자 분석시약의 성능 및 품질관리용 Plasma Working Standards 제조에 관한 연구)

  • Kim, Myung Han;Cho, Youn Jung;Kwon, So-Yong;Cho, Nam Sun
    • The Korean Journal of Blood Transfusion
    • /
    • v.23 no.2
    • /
    • pp.152-161
    • /
    • 2012
  • Background: Since Jan. 2012, for performance evaluation of viral reagents, analysis of domestic samples has been recommended in order to obtain approval from the KFDA when they are first introduced to Korea. This regulation requires the standard domestic materials driven from locally infected samples. We tried manufacturing the plasma working standards of HBV, HCV, and HIV NAT using a mixed titer of viral loads. Methods: Forty three HBV DNA positive plasmas, 25 HCV RNA positive plasmas, and 26 HIV RNA positive plasmas were evaluated according to viral load and genotype. Several plasma units, which had high-titer viral loads and the common viral genotypes in Korea, were selected as the source materials for each viral standard. To adjust the appropriate concentration based on the detectable range of variable viral reagents, the source plasma was diluted to several concentrations, divided into small vials, and analyzed for quantification. Results: The 13 plasma working standards, which had variable viral loads for the mixed titer performance panel of HIV, HCV, and HBV NAT, were produced. Conclusion: These national standard materials were first produced in order to supply the mixed titer performance panel for the viral NAT reagent of the level IV transfusion related high-risk group in Korea.

vfr, A Global Regulatory Gene, is Required for Pyrrolnitrin but not for Phenazine-1-carboxylic Acid Biosynthesis in Pseudomonas chlororaphis G05

  • Wu, Xia;Chi, Xiaoyan;Wang, Yanhua;Zhang, Kailu;Kai, Le;He, Qiuning;Tang, Jinxiu;Wang, Kewen;Sun, Longshuo;Hao, Xiuying;Xie, Weihai;Ge, Yihe
    • The Plant Pathology Journal
    • /
    • v.35 no.4
    • /
    • pp.351-361
    • /
    • 2019
  • In our previous study, pyrrolnitrin produced in Pseudomonas chlororaphis G05 plays more critical role in suppression of mycelial growth of some fungal pathogens that cause plant diseases in agriculture. Although some regulators for pyrrolnitrin biosynthesis were identified, the pyrrolnitrin regulation pathway was not fully constructed. During our screening novel regulator candidates, we obtained a white conjugant G05W02 while transposon mutagenesis was carried out between a fusion mutant $G05{\Delta}phz{\Delta}prn::lacZ$ and E. coli S17-1 (pUT/mini-Tn5Kan). By cloning and sequencing of the transposon-flanking DNA fragment, we found that a vfr gene in the conjugant G05W02 was disrupted with mini-Tn5Kan. In one other previous study on P. fluorescens, however, it was reported that the deletion of the vfr caused increased production of pyrrolnitrin and other antifungal metabolites. To confirm its regulatory function, we constructed the vfr-knockout mutant $G05{\Delta}vfr$ and $G05{\Delta}phz{\Delta}prn::lacZ{\Delta}vfr$. By quantifying ${\beta}-galactosidase$ activities, we found that deletion of the vfr decreased the prn operon expression dramatically. Meanwhile, by quantifying pyrrolnitrin production in the mutant $G05{\Delta}vfr$, we found that deficiency of the Vfr caused decreased pyrrolnitrin production. However, production of phenazine-1-carboxylic acid was same to that in the wild-type strain G05. Taken together, Vfr is required for pyrrolnitrin but not for phenazine-1-carboxylic acid biosynthesis in P. chlororaphis G05.

Identification and Characterization of Microbial Community in the Coelomic Fluid of Earthworm (Aporrectodea molleri)

  • Yakkou, Lamia;Houida, Sofia;Dominguez, Jorge;Raouane, Mohammed;Amghar, Souad;Harti, Abdellatif El
    • Microbiology and Biotechnology Letters
    • /
    • v.49 no.3
    • /
    • pp.391-402
    • /
    • 2021
  • Earthworms play an important role in soil fertilization, interacting continually with microorganisms. This study aims to demonstrate the existence of beneficial microorganisms living in the earthworm's immune system, the coelomic fluid. To achieve this goal, a molecular identification technique was performed, using cytochrome c oxidase I (COI) barcoding to identify abundant endogenic earthworms inhabiting the temperate zone of Rabat, Morocco. Then, 16S rDNA and ITS sequencing techniques were adopted for bacteria and fungi, respectively. Biochemical analysis, showed the ability of bacteria to produce characteristic enzymes and utilize substrates. Qualitative screening of plant growth-promoting traits, including nitrogen fixation, phosphate and potassium solubilization, and indole acetic acid (IAA) production, was also performed. The result of mitochondrial COI barcoding allowed the identification of the earthworm species Aporrectodea molleri. Phenotypic and genotypic studies of the sixteen isolated bacteria and the two isolated fungi showed that they belong to the Pseudomonas, Aeromonas, Bacillus, Buttiauxella, Enterobacter, Pantoea, and Raoultella, and the Penicillium genera, respectively. Most of the isolated bacteria in the coelomic fluid showed the ability to produce β-glucosidase, β-glucosaminidase, Glutamyl-β-naphthylamidase, and aminopeptidase enzymes, utilizing substrates like aliphatic thiol, sorbitol, and fatty acid ester. Furthermore, three bacteria were able to fix nitrogen, solubilize phosphate and potassium, and produce IAA. This initial study demonstrated that despite the immune property of earthworms' coelomic fluid, it harbors beneficial microorganisms. Thus, the presence of resistant microorganisms in the earthworm's immune system highlights a possible selection process at the coelomic fluid level.

Transcriptomic Analysis of the Difference of Bovine Satellite Cell Between Longissimus dorsi and Semimembranosus on Hanwoo Muscle Tissues (한우의 등심과 사태조직 유래 근육위성세포의 성장단계별 유전발현 차이 분석)

  • Kim, H.J.;Kang, D.H.;Park, B.H.;Lee, W.Y.;Choi, J.H.;Chung, K.Y.
    • Journal of Practical Agriculture & Fisheries Research
    • /
    • v.23 no.1
    • /
    • pp.117-128
    • /
    • 2021
  • The skeletal muscle development of Hanwoo steer has been processed in the prenatal and postnatal periods. Bovine satellite cell located in perimysium of muscle tissues has differentially distributed in peripheral tissues. The study of postnatal development of satellite cells can help understand the genetic and functional regulation of meat characteristics. Factors affecting muscle size increase are related to the accumulation of DNA or synthesis of RNA proteins. In this study, we observed muscle development and differentiation after culturing bovine satellite cells derived from longissimus dorsi and semimembranosus regions of Hanwoo muscle tissue. In addition, RNA sequencing data were analyzed for differentially expressed genes (DEG) involved in intracellular muscle development and growth. The DEG of the two muscle tissues were compared according to 1day, 2day, 4day, and 7day. The overall gene expression level was confirmed by the heat map. Gene Ontology (GO) classification method was used to compare the expression level of gene groups affecting LD and SM development. The histology of GO was consistent with the time-cause change of LD and SM cell morphology. SM showed more active skeletal muscle development than LD. Even within the same time, SM expressed more genes than LD, thus synthesizing more muscle fibers

The Philippines Coconut Genomics Initiatives: Updates and Opportunities for Capacity Building and Genomics Research Collaboration

  • Hayde Flandez-Galvez;Darlon V. Lantican;Anand Noel C. Manohar;Maria Luz J. Sison;Roanne R. Gardoce;Barbara L. Caoili;Alma O. Canama-Salinas;Melvin P. Dancel;Romnick A. Latina;Cris Q. Cortaga;Don Serville R. Reynoso;Michelle S. Guerrero;Susan M. Rivera;Ernesto E. Emmanuel;Cristeta Cueto;Consorcia E. Reano;Ramon L. Rivera;Don Emanuel M. Cardona;Edward Cedrick J. Fernandez ;Robert Patrick M. Cabangbang;Maria Salve C. Vasquez;Jomari C. Domingo;Reina Esther S. Caro;Alissa Carol M. Ibarra;Frenzee Kroeizha L. Pammit;Jen Daine L. Nocum;Angelica Kate G. Gumpal;Jesmar Cagayan;Ronilo M. Bajaro;Joseph P. Lagman;Cynthia R. Gulay;Noe Fernandez-Pozo;Susan R. Strickler;Lukas A. Mueller
    • Proceedings of the Korean Society of Crop Science Conference
    • /
    • 2022.10a
    • /
    • pp.30-30
    • /
    • 2022
  • Philippines is the second world supplier of coconut by-products. As its first major genomics project, the Philippine Genome Center program for Agriculture (PGC-Agriculture) took the challenge to sequence and assemble the whole coconut genome. The project aims to provide advance genetics tools for our collaborating coconut researchers while taking the opportunity to initiate local capacity. Combination of different NGS platforms was explored and the Philippine 'Catigan Green Dwarf' (CATD) variety was selected with the breeders to be the crop's reference genome. A high quality genome assembly of CATD was generated and used to characterize important genes of coconut towards the development of resilient and outstanding varieties especially for added high-value traits. The talk will present the significant results of the project as published in various papers including the first report of whole genome sequence of a dwarf coconut variety. Updates will include the challenges hurdled and specific applications such as gene mining for host insect resistance and screening for least damaged coconuts (thus potentially insect resistant varieties). Genome-wide DNA markers as published and genes related to coconut oil qualitative/quantitative traits will also be presented, including initial molecular/biochemical studies that support nutritional and medicinal claims. A web-based genome database is currently built for ease access and wider utility of these genomics tools. Indeed, a major milestone accomplished by the coconut genomics research team, which was facilitated with the all-out government support and strong collaboration among multidisciplinary experts and partnership with advance research institutes.

  • PDF

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.

Production of Colchicine Induced Tetraploids in Rye (Secale cereale L.) (콜히친 처리에 의한 4배체 호밀 육성)

  • Hwang, Jong-Jin;Kim, Dea-Wook;Kim, Chul-Woo;Son, Beum-Young;Baek, Seong-Beum;Park, Hyong-Ho;Ku, Ja-Hwan;Kim, Jung-Tae;Lee, Jin-Seok;Moon, Jung-Kyung;Kwon, Young-Up;Han, Ouk-Kyu
    • KOREAN JOURNAL OF CROP SCIENCE
    • /
    • v.57 no.4
    • /
    • pp.343-352
    • /
    • 2012
  • This experiment was carried out to report some data such as survival rate, tetraploid production efficiency, and agronomic characteristics of offspring from the induced tetraploid by the colchicine treatment in rye. The colchicine was soaked with 0.05%, 12 hours in dark condition and at two growth stages (green seed and 2nd leaf stage) in diploid ryes. Flow cytometry (FC) was proved to be efficient and rapid tool for screening ploidy levels in rye, showing around 40 to 60 in DNA amount (DAP1) corresponding to diploid and 80 to 110 tetraploid. There were 18.5% of survival rate at green seed treatment and 78% at 2nd leaf stage in average of two rye cultivars, Gogu and Jogreen, but in reverse 50.9% and 1.1% in the ratio of tetraploid to total tillers among the plants survived, respectively, resulting in 9.42% of tetraploid production rate in green seed treatment and 0.86% at 2nd leaf stage, respectively. In green seed treatment, there were 33% of survival rate in Gogu, 4% in Jogreen in 1st year, but 56% in Gogu, 21% in Jogreen and 49% in Charmgreen, respectively. The rate of tetraploid to total spikes among survived was 53.7% in Gogu, 32.4% in Jogreen, and 50.9% in average in 1st year, and 64.1% in Gogu, 51.5% in Jogreen, 60% in Charmgreen, and 60.5% in average in 2nd year. In green seed treatment, tetraploid production rate (survival rate ${\times}$ tetraploid ratio ${\times}$ 100) was 17.7% in Gogu and 1.3% in Jogreen and 9.42% in average in 1st year, and 35.9% in Gogu, 10.8% in Jogreen, 29.4% in Charmgreen, and 25.4% in average of three diploid rye cultivars. By the colchicine treatment with 0.05% for 12 hours in Gogu and Jogreen, 35 tetraploid plants were obtained and they produced 2,673 seeds with 148 spikes. There were 3.3-4.4 in the number of spikes per plant, 15.6-18.3 in grain number per spike, and 37.6 g in Gogu and 46.8 g in Jogreen in the 1,000-grain weight.