• Title/Summary/Keyword: rDNA sequencing

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Association Study Between the Polymorphisms of Exostosin-1 Gene and Economic Traits in Hanwoo (한우 Exostosin-1 유전자의 SNP 탐색 및 경제형질 관련성 분석)

  • Kim, Bum-Soo;Kim, Nam-Kuk;Lee, Seung-Hwan;Cho, Yong-Min;Heo, Kang-Nyeong;Park, Eung-Woo;Yang, Boo-Keun;Yoon, Du-Hak
    • Journal of Animal Science and Technology
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    • v.53 no.1
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    • pp.7-13
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    • 2011
  • The aim of this study was to identify the polymorphism on exostosin-1 (EXT1) gene and to associate with economic traits in Hanwoo (Korean cattle). We sequenced for detection of single nucleotide polymorphism (SNP) with 24 unrelated individuals and identified four SNPs (T272196A, C272359T, G290964A and A302092G). Relationship between the genotypes of 583 Hanwoo individuals by PCR-RFLP and economic traits were analyzed by general linear model. In EXT1 gene, there were four SNPs associated with economic traits such as eye muscle area breeding value, marbling score breeding value, backfat and thickness breeding value (p<0.05 to p<0.01). In conclusion, this study indicates an important role of EXT1 gene in determining the meat quality or economic characteristics in Hanwoo.

Characterization of the cloned RNA1 gene of Saccharomyces cerevisiae (Cloning된 효모의 RNAI 유전자의 특성에 관하여)

  • Song, Young-Hwan;Kim, Dae-Young;Kim, Jin-Kyung
    • Journal of fish pathology
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    • v.6 no.2
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    • pp.93-101
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    • 1993
  • The RNAI mutation of Saccharomyces cerevisia is a recessive and temperature sensitive lethal mutation which interferes with the production of mRNA, rRNA, and tRNA. However, the precise role of RNAI gene have not been revealed until yet. We have cloned rna1-1 mutant gene from rna1-1 mutant yeast strain(R49 ; trpl, ura3-52, rna1-1). The 3.4kb BglII fragment of wild type RNAI clone(81-2-6) contains whole RNAI gene. The genomic southern blotting with BglII digested R49 genomic DNA as a probe shows the unique and identical band with wild type 3.4kb BglII fragment. Therefore, We prepared partial BglII genomic library(3~4kb BglII fragments) into BamH I site of pUC19. The rna 1-1 mutant clone was screened with Digoxigenin(DIG)-lableled probe by high density colony hybridization. The 5'-flanking region of rna1-1 gene was sequenced by dideoxy chain termination method. The 5'-flanking sequence of RNAI gene contains three TATA-like sequence ; TAATA, TATA and TTTTAA at position of -67, -45, and -36 from first ATG codon respectively. The 5'-flanking region of wild type RNA I gene from ATG codon to -103nt was deleted with Bal31 exonuclease digestion, generating $pUC{\Delta}$/RNA I. After constructing $pYEP{\Delta}RNA$ I (consists of -103nt deleting RNA I gene, URA3 gene, $2{\mu}m$ rep. origin), pYEPrna1-1(consists of Xba I fragment of pUCrna1-1. URA3 gene, $2{\mu}m$ rep. origin), and pYEPRNAI. each plasmid was transformed into host strain(trpl, ura3-52, rna1-1) by electroporation, respectively. Yeast transformant carrying $pYEP{\Delta}RNA$ I did not complement the thermal sensitivity of rna1-1 gene. It means that TATA-like sequences in 5'-flanking region is not TATA sequence for transcribing RNAI gene and there may be other essential sequence in upstream region for the transcription of RNAI gene.

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Genetic Variations of Chicken MC1R Gene and Associations with Feather Color of Korean Native Chicken (KNC) 'Woorimatdag' (토종 '우리맛닭' 부계 및 실용계에서 MC1R 유전자 변이 및 모색과의 연관성 분석)

  • Park, Mi Na;Kim, Tae-Hun;Lee, Hyun-Jeong;Choi, Jin Ae;Heo, Kang-Nyeong;Kim, Chong-Dae;Choo, Hyo-Jun;Han, Jae-Yong;Lee, Taeheon;Lee, Jun-Heon;Lee, Kyung-Tai
    • Korean Journal of Poultry Science
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    • v.40 no.2
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    • pp.139-145
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    • 2013
  • There are several loci controlling the feather color of birds, of which one of the most studied is Extended black (E) encoding the melanocortin 1-receptor (MC1R). Mutations in this gene affect the relative distribution of eumelanin, phaeomelanin. The association of feather color and sequence polymorphism in the melanocortin 1-receptor (MC1R) gene was investigated using Korean native chicken H breed (H_PL) and 'Woorimatdag' commercial chickens (Woorimatdag_CC). In order to correlate gene mutation to Korean native chicken feather color, single nucleotide polymorphism (SNP) from MC1R gene sequence were investigated. A total of 307 birds from H_PL and Woorimatdag_CC were used. H_PL have black, black-brown feather color and Woorimatdag_CC have black with brown spots or brown with black spots. There are 6 SNPs in MC1R gene, locus T69C, C212T, A274G, G376A, G636A, T637C. 3 SNPs are nonsynonymous that change amino acid. But it is difficult to find correlation of feather color and polymorphisms. It will be needed to increase the population of Korean native chicken H breed and correlation analysis of genetic variation with feather colors.

Characterization of Noble AmpC-Type $\beta$-Lactamases Among Clinical Isolates Using New Expression/Secretion Vector (발현ㆍ분비 벡터 및 임상 균주가 생성하는 신규 AmpC-type $\beta$-lactamase의 특성)

  • 정하일;성광훈;이정훈;장선주;이상희
    • Korean Journal of Microbiology
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    • v.40 no.2
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    • pp.104-110
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    • 2004
  • To determine evolution and genotype of new chromosomal AmpC $\beta$-lactamases among clinical isolates of Enterobacter species, we performed antibiotic susceptibility testing, pI determination, sequencing, and phy-logenetic analysis using developed expression/secretion vector. Six isolates have shown to produce AmpC $\beta$-lactamases. Six genes of AmpC $\beta$-lactamases that are responsible for the resistance to cephamycins (cefoxitin and cefotetan), amoxicillin, cephalothin, and amoxicillin-clavulanic acid were cloned and characterized in pMSG12119. Insert fragment containing the ampC genes was sequenced and found to have an open reading frame coding for 381-amino-acid $\beta$-lactamase. The nucleotide sequence of four ampC genes ($bla_EcloK992004.l$, $bla_EcloK995120.1$, $bla_EcloK99230$, and $bla_EareK9911729$) shared considerable homology with that of chromosomal ampC gene ($bla_EcloMHN1$) of E. cloacae MHN1 (more than 99.6% identity). The sequences of two ampC genes ($bla_EcloK9973$ and $bla_EcloK9914325$) showed close similarity to the chromosomal ampC gene ($bla_EcloQ908R$) of E. clo-acae 908R (99.7% identity). The results from phylogenetic analysis suggested that six ampC genes could be originated from $bla_EcloMHN1$ / or $bla_EcloQ908R$ / MIC patterns and exact pI values of six transformants indicated that the developed expression/secretion vector (pMSG1219) was suitable for the characterization of foreign genes in E. coli strain.

Helicobacter pylori vacA Mosaicism and New Primers for vacA Signal Sequence Indigenous to Korea (Helicobacter pylori vacA 대립유전자의 Mosaicism과 Signal Sequence의 한국고유 시발체)

  • Ahn, Yeon-Hwa;Kim, Heung-Ryel;Lee, Ji-Eun;Hwang, Tae-Sook;Choe, Yon-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.4 no.2
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    • pp.155-160
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    • 2001
  • Purpose: Helicobacter pylori has been known to have diverse vacA allelic types. The purpose of the study was to identify vacA diversity in Korea and design new primers for signal sequence alleles indigenous to Korea. Methods: Fifty antral biopsy specimens, which had been proven to be H. pylori-positive, were examined for vacA status; signal sequence and mid-region. After PCR amplification and DNA sequencing, vacA alleles of Korean H. pylori strains were compared with those from other countries. Results: Among Korean H. pylori strains vacA alleles with all combinations of signal sequence and mid-region were found, with the exception of s1b or s2. vacA genotype s1c/m1 was predominant in Korea. We found that GGGAGCGTTR in s1a and GGGGYTATTG in s1c were the indigenous sequences to Korea and constructed the new Korean specific primers for the vacA signal sequence; VASK-F, VASK-R, S1AK-F, and S1CK-F. Conclusion: This study showed that s1c/m1 is the predominant type of vacA allele in Korea. We designed new primers for the vacA signal sequence.

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Identification of a Newly Isolated Protease-producing Bacterium, Bacillus subtilis FBL-1, from Soil (토양으로부터 새로이 분리된 단백질 분해효소 생산 미생물 Bacillus subtilis FBL-1의 동정)

  • Kim, Mina;Si, Jin-Beom;Wee, Young-Jung
    • Microbiology and Biotechnology Letters
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    • v.44 no.2
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    • pp.185-193
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    • 2016
  • A novel proteolytic bacterium was isolated from soil at Yeungnam University, South Korea. The strain, named FBL-1, was rod-shaped with a smooth surface. Biolog and API 50CHB test results revealed that strain FBL-1 was a Bacillus species. Based on 16S rDNA sequencing and chemotaxonomic characterization, the strain was identified as Bacillus subtilis because it had the highest homology with Bacillus subtilis subsp. subtilis NCIB 3610 (99.5%). In liquid culture at 37℃ with shaking at 200 rpm, fructose and yeast extract were found to be the best carbon and nitrogen sources, respectively, for cell growth and protease production. The highest protease activity (451.640 U/ml) was obtained when the strain was cultured in medium containing 20 g/l of fructose and 5 g/l of yeast extract. Although further studies are needed to characterize the protease and enhance its activity, the newly isolated protein-degrading B. subtilis FBL-1 can be applicable for the production of peptides and for the degradation of proteins in various industries.

Identification and Cultural Characterization of Antioxidant Producing Bacteria Isolated from the Jeju Coasts (제주 연안에서 분리한 항산화물질을 생산하는 균주의 동정 및 배양학적 특성)

  • Kim Man-Chul;Heo Moon-Soo
    • Journal of Life Science
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    • v.15 no.5 s.72
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    • pp.749-754
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    • 2005
  • An antioxidant- producing bacterium was isolated from sea water in Jeju island. The isolated strain, SC2-1 was gram-positive, catalase positive, facultatively anaerobic, oxidase negative, motile and small rods. The strain utilized sucrose, dextrose, fructose, mannitol and maltose as a sole carbon and energy source and sodium chloride was required for the bacteria growth. The radical scavenging activity of the culture supernatants was determined by DPPH (1,1-diphenyl-2-picrylhydrazyl) method. This bacterium was identified based on cellular fatty acids analysis and 16S rDNA sequencing, and then named Exiguobacterium sp. SC2-1. The modified optimal medium compositions required the addition of maltose $2.5\%(w/v)$, yeast extract $1.5\%(w/v)$ and $KH_{2} PO_{4} 0.05\%(w/v)$ in marine broth (Difco. Co. USA). Antioxidant activity of under optimal culture conditions were $93\%$.

Induced of Systemic Resistance against Gray Leaf Spot in Pepper by Enterobacter Species Isolated from Family Gramineae Plants in Dok-do (독도의 벼과식물로부터 분리된 Enterobacter spp.에 의한 고추의 흰별무늬병에 대한 전신유도저항성)

  • Son, Jin-Soo;Sumayo, Marilyn;Kang, Hyun-Uk;Kim, Byung-Soo;Kwon, Duck-Kee;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.40 no.2
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    • pp.135-143
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    • 2012
  • This study's aim is to isolate and characterize plant growth promoting Enterobacter species for the biological control of gray leaf spot in pepper. Screening was carried out from the rhizosphere of Agropyron tsukushiensi var. transiens (Hack.) Ohwi in Dok-do. Rhizobacterial isolates were partially identified by 16S rDNA sequencing and Enterobacter species were tested for plant growth promoting capabilities and the induction of systemic resistance in pepper against gray leaf spot caused by Stemphylium solani. Isolates were tested for production of indole-acetic acid and siderophore, and for phosphate solubilization. The application of isolates was effective in controlling gray leaf spot in pepper with E. asburiae (KNUC5007) and E. cancerogenes (KNUC5008 and KNUC5010) having the highest efficacy in reducing gray leaf spot severity. This is the first report of the biological control of gray leaf spot in pepper using rhizobacteria and it is hoped that this study will increase the utilization of Enterobacter species as plant growth promoters and biocontrol agents.

Microbial Structure and Community of RBC Biofilm Removing Nitrate and Phosphorus from Domestic Wastewater

  • Lee, Han-Woong;Choi, Eui-So;Yun, Zu-Whan;Park, Yong-Keun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1459-1469
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    • 2008
  • Using a rotating biological contactor modified with a sequencing bath reactor system (SBRBC) designed and operated to remove phosphate and nitrogen [58], the microbial community structure of the biofilm from the SBRBC system was characterized based on the extracellular polymeric substance (EPS) constituents, electron microscopy, and molecular techniques. Protein and carbohydrate were identified as the major EPS constituents at three different biofilm thicknesses, where the amount of EPS and bacterial cell number were highest in the initial thickness of 0-100${\mu}m$. However, the percent of carbohydrate in the total amount of EPS decreased by about 11.23%, whereas the percent of protein increased by about 11.15% as the biofilm grew. Thus, an abundant quantity of EPS and cell mass, as well as a specific quality of EPS were apparently needed to attach to the substratum in the first step of the biofilm growth. A FISH analysis revealed that the dominant phylogenetic group was $\beta$- and $\gamma$-Proteobacteria, where a significant subclass of Proteobacteria for removing phosphate and/or nitrate was found within a biofilm thickness of 0-250${\mu}m$. In addition, 16S rDNA clone libraries revealed that Klebsiella sp. and Citrobacter sp. were most dominant within the initial biofilm thickness of 0-250${\mu}m$, whereas sulfur-oxidizing bacteria, such as Beggiatoa sp. and Thiothrix sp., were detected in a biofilm thickness over 250${\mu}m$. The results of the bacterial community structure analysis using molecular techniques agreed with the results of the morphological structure based on scanning electron microscopy. Therefore, the overall results indicated that coliform bacteria participated in the nitrate and phosphorus removal when using the SBRBC system. Moreover, the structure of the biofilm was also found to be related to the EPS constituents, as well as the nitrogen and phosphate removal efficiency. Consequently, since this is the first identification of the bacterial community and structure of the biofilm from an RBC simultaneously removing nitrogen and phosphate from domestic wastewater, and it is hoped that the present results may provide a foundation for understanding nitrate and phosphate removal by an RBC system.

A Study on the TCE/PCE Removal Using Biofiltration and the Microbial Communities Variation Using DGGE Method (생물 여과를 이용한 TCE/PCE제거 및 DGGE법을 이용한 관련미생물 군집변화에 관한 연구)

  • Kim, Eung-In;Park, Ok-Hyun;Jung, In-Gyung
    • Journal of Korean Society of Environmental Engineers
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    • v.30 no.11
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    • pp.1161-1169
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    • 2008
  • The removals of TCE and PCE vapor with or without a supply of toluene as a primary substrate were compared in a biofiltration process, and the variations of microbial communities associated with the removal were also investigated. As a result of investigations on the removals of TCE/PCE in a biofilter B within which TCE/PCE-acclimated sludge was attached on the surface of media without a supply of primary substrate, and those in another biofilter A where toluene-acclimated sludge was attached with a supply of toluene as a primary substrate, followings were found: (i) parts of microbes responsible to the decomposition of toluene vapor participate in the removal of chlorinated VOCs such as TCE and PCE, and (ii) effective biological removals of TCE and PCE vapor do not necessarily need cometabolism. Sequencing of 16S rDNA obtained from the band profile of DGGE (Denaturating Gradient Gel Electrophoresis), it was confirmed that: (i) uncultured alpha proteobacterium, uncultured Desulfitobacterium, uncultured Rhodobacteraceae bacterium, Cupriavidus necator, and Pseudomonas putida were found to be toluene-decomposing microbes, (ii) alpha proteobacterium HTCC396 is a TCE-removing microbe, (iii) Desulfitobacterium sp. is a PCE-decomposing microbe, and (iv) particularly, uncultured Desulfitobacterium sp. is probably a microbe decomposable not only toluene but also various chlorinated VOC vapor including TCE and PCE.