• Title/Summary/Keyword: DNA code

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Cloning and Expression of the metE gene coding for homocysteine methyltransferase from the basidiomycete Ganoderma lucidum in E. coli (영지버섯으로부터 homocysteine methyltransferase를 암호화 하는 metE 유전자의 클로닝 및 E. coli에서의 발현)

  • Kim, Hyun-Jeong;Park, Dong-Chul;Lee, Kap-Duk;Lee, Byul-La;Lee, Kap-Rang
    • The Korean Journal of Mycology
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    • v.21 no.4
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    • pp.279-284
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    • 1993
  • The metE gene coding for $N^{5}-methyl-H_{4}-folate;$ homocysteine methyltransferase from the basidiomycete Ganoderma lucidum was cloned by complementation of methionine-requiring mutants of E. coli. The size of a inserted DNA was about 1.54 kb and had 5 restriction enzyme sites. A physical map was constructed. Southern blot analysis confirmed the presence of a transforming DNA in the genome of Ganoderma lucidum. indicating the presence of a single copy.

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Nucleotide sequence analysis of a second set of the polyketide synthase .betha.-ketoacyl synthase and chain length factor genes from the salinomycin-producing streptomyces albus

  • Hyun, Chang-Gu;Park, Kwan-Hyung;C.Richard Hutchinson;Suh, Joo-Won
    • Journal of Microbiology
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    • v.35 no.1
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    • pp.40-46
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    • 1997
  • The pWHM220 cosmid with a 24-kb insert cloned from Streptomyces albus ATCC 21838 induces the biosynthesis of a polysther antibiotic similar to salinomycin in Streptomyces invidans. We have analyzed this region by DNA sequencing as well as Southern blot hybridization with type I and type II polyketide synthase (PKS) probes. Surprisingly, we found another set of type II SKS genes only 10-kb from the original PKS genes, salABCDE. The DNA sequence revealed two complete open reading frames (ORFs) named salB2 and salC2, and one partial ORF that does not resemble any known DNA or deduced protein sequence. The salC2 should code for chain length determining factor while the deduced amino acid sequence encoded by salB2 exhibits high similarity to .betha.-ketoacyl synthase from different PKS gene clusters. The highest identity was found for .betha.-keetoacyl synthases from S. argillaceus (MtmP. 59.1% identity), the mithramycin producer and from S. venezuelae ISP5230 (JadA, 52.3% identity), the jadomycin producer. The SalC2 protein clearly resembles its counterparts in order aromatic PKS gene clusters that are believed to influence the length of the polyketide chain. The highest identities observed were to that of S. argillaceus (MtmK, 62.3%) and S. venezuelae ISP 5230 (JadB, 55.1%) proteins, Moreover, the deduced amino acid sequences of the salB2 and salC2 products were 29.0% identical.

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Effects of 835-MHz Radiofrequency Radiation on the Chromosomal DNA of Mouse Thymic Lymphoma L5178Y $Tk^{+/-}$ Cells

  • Choi, Jong-Soon;Son, TaeHo;Chang, Sung-Keun;Hong, Sae-Yong
    • Korean Journal of Environmental Biology
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    • v.22 no.4
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    • pp.507-512
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    • 2004
  • This study was focused on the risk assessment of whether radiofrequency electromagnetic fields generated by mobile phone is cytogenetically toxic or not. We conducted the effects of 835-MHz electromagnetic field (EMF) on DNA strand breaks in mouse thymic lymphoma L5178Y $Tk^{+/1-}$ cells using alkaline comet assay. EMF frequency 835-MHz we chosen is one of the most popular communication frequency bands in Korean code-division multiple-access (CDMA) mobile phone system. The cells were exposed to 835-MHz EMF alone or 835-MHz EMF combined with cyclophosamide(CPA) or 4-nitroquinoline-1-oxide (4NQO) at specific absorption rate (SAR) of 4.0 W $kg^{-l}$ for 24 and 48hrs. DNA damage expressed as tail moment was increased more than two-fold after exposure to 835-MHz EMF for 24 and 48hr. In particular, CPA for 48hr and 4NQO for 24 hr enhanced notably the tail moment to 9-fold and 16-fold in the presence of 835-MHz EMF, respectively, compared to each single treatment. From these results, it appears that exposure to CDMA-mobile phone radiation at 835-MHz frequency may potentiate DNA strand breaks of mouse thymic lymphoma L5178Y $Tk^{+/1-}$;cells under the defined conditions of this study.

Application of DNA Test for Individual Traceability in the Brand Marketing of Korean Native Pig. (한국 재래돼지 브랜드 돈육 원산지 검증을 위한 유전자 원산지 감식 기법 활용 연구)

  • Choi, Bong-Am;Lee, Hak-Kyo;Jeon, Gwang-Joo;Oh, Jaen-Don;Choi, Il-Sin;Park, Mi-Hyun;Kong, Hong-Sik;Jung, Il-Jung;Kim, Tae-Hun;Yoon, Doo-Hak;Cho, Byung-Wook
    • Korean Journal of Organic Agriculture
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    • v.12 no.2
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    • pp.197-207
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    • 2004
  • Identification of animals has been used with an e ar tag with dummy code and blood typing has been used for paternity and individual identification in live animals. Various genetic markers are different for breeds of pig and hence, it is necessary to identity the discrete genetic marker in korean native pig. A total of 240 pigs were used to find korean native pig population specific markers that expressed in population of korean native pigs. To identify the individual traceability, 20 animals were randomly chosen and tested for a whole process from being live to slaughter stages. The candidate genetic marker used in the study were 18 DNA microsatellites which were identified in pig genome. The number of alleles of those DNA microsatellites ranged form a minimum of 3 to maximum of 6. The heterozygote frequency rang6d from 0.44 to 0.69. Effective number of alleles for each DNA microsatellotes were 2 to 4. By choosing 6 candidate genetic markers among all, the traceability of individual identification was estimated as accurate as 99.99%(p>0.0014), nearly.

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Food Fraud Monitoring of Raw Materials for Commercial Seafood Products Using DNA Barcode Information (DNA Barcode를 이용한 수산가공품 원재료 진위판별)

  • Park, Eun-Ji;Kang, Ju-Yeong;Lee, Han-Cheol;Park, Min-Ji;Yang, Ji-Young;Shin, Ji-Young;Kim, Gun-Do;Kim, Jong-Oh;Seo, Yong-Bae;Kim, Jung-Beom
    • Journal of Food Hygiene and Safety
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    • v.36 no.4
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    • pp.331-341
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    • 2021
  • DNA barcode sequences of commercial seafood products, which are difficult to morphologically discriminate, were analyzed to determine cases of food fraud. The gene sequences were analyzed by amplifying the COX I (cytochrome C oxidase subunit I) gene region of mitochondrial DNA, which is mainly used for species identification. The DNA barcode sequences were compared with the gene sequence of each fish registered in the US National Center for Biotechnology. A total of 46 processed seafood products (12 Pagrus majo, 4 Oplegnathus fasciatus, 7 Dentex tumifrons, 2 Acanthopagrus schlegelii, 7 Oreochromis niloticus, 6 Branchiostegus japonicus, 8 Branchiostegus albus) were investigated. Having DNA sequence identity of more than 97% was judged as the same species. As a result of this study, no cases of forgery and alteration were detected. However, some disparities in the commercial names used in local markets and the standard names given in the Korea Food Code were found, which may cause confusion for consumers. It is therefore suggested that the standard name or scientific name be displayed on seafood product labels.

Moleculay Cloning of the cDNA Encoding the 16 kDa Subunit of V-ATPase in Rat Brain (흰쥐 뇌에서 발현되는 16 kDa Vacuolar (H$^{+}$)-ATPase의 유전자 클로닝)

  • Shin, Song-Woo;Yoo, Min
    • Biomedical Science Letters
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    • v.6 no.3
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    • pp.165-170
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    • 2000
  • Vacuolar (H$^{+}$)-ATPase (V-ATPase) is an intracellular protein which consists of multiple subunits. It carries out acidification by pumping protons in the cell. This enzyme has also been found in the synaptic vesicles and may play an important role in the neurotransmission. We cloned cDNA fragments encoding the 16 kDa subunit of V-ATPase from the rat brain by RT-PCR and PCR using total RNA or recombinant phage DNA as templates. They contained the full coding sequences (468 bp) and one nucleotide at 3' region turned out to be different (A to C) when compared to the liver counterpart. However, this polymorphic difference did not cause any significant change in the primary structure of the protein because both GCA and GCC code for alanine. Our study would contribute to the understanding of the function of 16 M)a V-ATPase in the brain and of the mechanisms of neurotransmission.

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A Study on Sasang Constitutional Gene Selection Using DNA Chips by Multivariate Analysis (유전자 칩 및 다변량 분석방법을 이용한 사상체질 유전자 선별에 관한 연구)

  • Kim, Pan-Joon;Seo, Eun-Hee;Lee, Jung-Hwan;Ha, Jin-Ho;Choi, Hong-Sik;Jung, Tae-Young;Goo, Deok-Mo
    • Journal of Sasang Constitutional Medicine
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    • v.18 no.3
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    • pp.131-144
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    • 2006
  • 1. Objectives This research uses the DNA chip, which includes 16,383 gene code, and various statistic prediction way that shows objectification index for the objectification of constitution diagnosis. 2. Methods Drawing blood whose constitution is confirmed, and analyze its gene information by using 1.7k DNA chip to find the gene correlation through multivariate statistical method. 3. Results and Conclusions Distinctive genes such as AK001919, U09384, NM_001805, X99962, NM_004796, AK026738, AL050148, BC002538, AK027074, AK026219, AF087962, AL390142, NM_015372, AL157466, NM_002446, AK024523, NM_014706, NM_014746 and AL137544 were related to Taeumin; AL157448, NM_005957, NM_005656, NM_017548, AK027246, NM_003025, NM_012302 and NM_005905 were represented in Soeumin, while AK026503, AF147325, NM_002076, AF147307, AK001375, NM_003740, NM_005114, AB007890, NM_005505, NM_015900, NM_014936, Z70694, AB023154, U52076, NM_004360, NM_005835, NM_017528, AF087987, NM_014897, AK021720, NM_006420, AJ277915, AK002118 and AK021918 were for Soyangin. This study figured out the possibility to develop the prediction system by sorting each constitution's gene, and research each constitution's distinctive character of manifestation pattern.

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Application of DNA Test for Individual Traceability in Hanwoo (Korean Cattle) (한우의 개체 추적 검증을 위한 유전자 감식 기법 활용 연구)

  • 이학교;전광주;공홍식;오재돈;최일신;김종대;조창연;윤두학;신형두
    • Food Science of Animal Resources
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    • v.24 no.1
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    • pp.8-14
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    • 2004
  • Identification of animals has been made with an ear tag with dummy code, and blood typing has been used for paternity and individual identification in live animals. As various genetic markers are for different cattle breeds vary, the discrete genetic markers are necessary to identify Hanwoo. A total of 740 progeny testing Hanwoo were used to identify Hanwoo specific markers. To examine traceability of individuals by using breed specific genetic codes, four animal were randomly sampled, and traced from live animals to post-slaughter processing stages. The candidate genetic makers used in the study were 16 DNA microsatellites which were identified in romosomes 1 and 14. The number of alleles of those DNA microsatellites ranged from a minimum of 3 to maximum of 12. The heterozygote frequency ranged from 0.022 to 0.824. Effective number of alleles for each DNA microsatellites were 3 to 6. Six selected candidate genetic markers were able ti trace individual cattle with an 100% confidence level.

Analysis of Bacterial Community Structure in the Soil and Root System by 168 rRNA Genes (16S rDNA를 이용한 토양, 작물근계의 세균군집 구조해석)

  • Kim, Jong-Shik;Kwon, Soon-Wo;Ryu, Jin-Chang;Yahng, Chang-Sool
    • Korean Journal of Soil Science and Fertilizer
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    • v.33 no.4
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    • pp.266-274
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    • 2000
  • Understanding of microbial community structure in soil-root system is necessary to use beneficial soil and rhizosphere microbes for improvement of crop production and biocontrol. The knowledge of behavior and function of microbes in soil-root system plays a key role for the application of beneficial inocula. Because the majority of the intact bacteria in soil are unable to grow on nutrient media, both culturable and nonculturable bacteria have to be studied together. In our study, culture-independent survey of bacterial community in the soil-root system of red pepper fields was conducted by the sequence analysis of three universal clone libraries of genes which code for small-subunit rRNA (rDNA). Universal small subunit rRNA primers were used to amplify DNA extracted from each sample and PCR products were cloned into pGEM-T. Out of 27 clones sequenced, 25 clones were from domain bacteria. Two of the rDNA sequences were derived from eukaryotic organelles. Within the domain bacteria, several kingdoms were represented : the Proteobacteria (16 clones). Cytophyga-Flexibacter-Bacteroides group (2 clones). the high G+C content gram-positive group(1 clone) and 4 unknown clones.

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Cloning of a ${\beta}-Xylosidase$ Gene from Alkalophilic Bacillus sp. and its Expression in Escherichia coli

  • Yu, Ju-Hyun;Kang, Yun-Sook;Park, Young-Seo
    • Journal of Microbiology and Biotechnology
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    • v.1 no.1
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    • pp.17-21
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    • 1991
  • A gene coding for ${\beta}-xylosidase$ in alkalophilic Bacillus sp. YC-335 isolated from soil was cloned into Escherichia coli HB101 using plasmid pBR322. The recombinant plasmid pYK40 was isolated, and the cloned HindIII fragment was 15 kilobases (kb). To reduce the size of the inserted DNA fragment of pYK40, the 15 kb HindIII fragment was subjected to a series of subclonings. A 6 kb subfragment was found to code for ${\beta}-xylosidase$ activity, and the recombinant plasmid was named pYK44. Southern hybridization analysis revealed that the cloned gene hybridized with 3.5 kb, 1.5 kb, and 1.0 kb of HindIII cleaved chromosomal DNA from Bacillus sp. YC-335. ${\beta}-xylosidase$ activity produced by recombinant E. coli was found to be 11 times higher than that produced by Bacillus sp. YC-335. Xylan was required to induce the production of ${\beta}-xylosidase$ in Bacillus sp. YC-335.

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