• Title/Summary/Keyword: specific DNA.

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Isolation and Characterization of a Pollen-specific cDNA Clone from Easter Lily

  • Kim, Seong-Ryong;An, Gyu-Heung
    • Journal of Plant Biology
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    • v.39 no.3
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    • pp.197-202
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    • 1996
  • A pollen-specific cDNA clone, LMP50, was isolated from the mature pollen cDNA library of the Easter lily. The LMP50 transcript was highly abundant in mture pollen grains but not detectable in other organs. The LMP50 cDNA clone contains 1383 nucleotides and two open reading frames. The first codes for a peptide of 15 amino acid residues. The role of this peptide is nuclear. The second encodes a protein containing 329 amino acid residues. This protein exhibited a significant homology to human tartrate-resistant acid phosphatase and porcine uteroferrin. Both of these enzymes have been suggested to play a role in iron transport. Therefore, LMP50 may act as an iron carrier protein in mature pollen grains.

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DNA Light-strand Preferential Recognition of Human Mitochondria Transcription Termination Factor mTERF

  • Nam, Sang-Chul;Kang, Chang-Won
    • BMB Reports
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    • v.38 no.6
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    • pp.690-694
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    • 2005
  • Transcription termination of the human mitochondrial genome requires specific binding to termination factor mTERF. In this study, mTERF was produced in E. coli and purified by two-step chromatography. mTERF-binding DNA sequences were isolated from a pool of randomized sequences by the repeated selection of bound sequences by gel-mobility shift assay and polymerase chain reaction. Sequencing and comparison of the 23 isolated clones revealed a 16-bp consensus sequence of 5'-GTG$\b{TGGC}$AGANCCNGG-3' in the light-strand (underlined residues were absolutely conserved), which nicely matched the genomic 13-bp terminator sequence 5'-$\b{TGGC}$AGAGCCCGG-3'. Moreover, mTERF binding assays of heteroduplex and single-stranded DNAs showed mTERF recognized the light strand in preference to the heavy strand. The preferential binding of mTERF with the light-strand may explain its distinct orientation-dependent termination activity.

Overproduction and Operator DNA-Protein Blotting of R100 Mutant MerR from Shigella flexneri

  • Yoon, Kyung-Pyo
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.250-255
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    • 1994
  • Wild-type and four mutant R100 merR genes were cloned and the proteins overproduced under tac promoter control of pKK223-3. His118Ala, Cys117Ser, Cys126Ser, and wild-type MerR were successfully overproduced although amino-terminal 14 amino acids deletion mutant MerR was not successful. The amount of overproduced wild-type MerR protein as well as other mutant MerR was between 15%-20% of the total protein. The protein was able to be purified up to 95% homogeneity. Specific DNA-protein blotting experiments showed that the 95 bp operator containing DNA fragment could bind to Cys126Ser, His118Ala, and wild- type MerR, but not to Cys117Ser. These results were consistent with the previously reported complementation experiment results that His118Ala, Cys126Ser, and wild-type MerR could repress the mer operon but Cys117Ser could not.

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Expression of Carboxypeptidase Z cDNA from Absidia zychae in saccharomyces cerevisiae and its characteristics (Saccharomyces cerevisiae에서 Absidia zychae 의 Carboxypeptidase A cDNA 의 발현과 특성)

  • 이병로;김종화
    • Microbiology and Biotechnology Letters
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    • v.23 no.2
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    • pp.150-155
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    • 1995
  • Carboxypeptidase Z(CPZ) cDNA of Absidia zychae was experssed in Saccharomyces cerevisiae. The expressed CPZ(YCPZ) was secreted about 30 mg/l into the medium and has a little higher molecular weight than the wild type CPZ in SDS-PAGE. By the result of N-terminal amino acid sequencing, YCPZ has additional 15 amino acids residues in N-terminus of CPZ. But YCPZ shows no difference with CPZ in enzyme activity and substrate specificity. For the identification of processing mechanism of YCPZ, 36-Arg was changed to 36-Thr by site specific mutagenesis. Mutant YCPZ does not processed at 36-Thr. It was, therefore, concluded that the YCPZ was processed by KEX2. According to endo F treatment, high amount of carbohydrate was N-glycosylated in YCPZ.

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Metagenomic Analysis of BTEX-Contaminated Forest Soil Microcosm

  • Ji, Sang-Chun;Kim, Doc-Kyu;Yoon, Jung-Hoon;Lee, Choong-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.17 no.4
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    • pp.668-672
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    • 2007
  • A microcosmal experiment using a metagenomic technique was designed to assess the effect of BTEX (benzene, toluene, ethylbenzene, and xylenes) on an indigenous bacterial community in a Daejeon forest soil. A compositional shift of bacterial groups in an artificial BTEX-contaminated soil was examined by the 16S rDNA PCR-DGGE method. Phylogenetic analysis of 16S rDNAs in the dominant DGGE bands showed that the number of Actinobacteria and Bacillus populations increased. To confirm these observations, we performed PCR to amplify the 23S rDNA and 16S rDNA against the sample metagenome using Actinobacteria-targeting and Bacilli-specific primer sets, respectively. The result further confirmed that a bacterial community containing Actinobacteria and Bacillus was affected by BTEX.

DNA Barcoding of Eurydice longiantennata (Isopoda, Cymothooidea, Cirolanidae) from South Korea

  • Kim, Sung Hoon;Choi, Hyun Ki;Kim, Jong Guk
    • Animal Systematics, Evolution and Diversity
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    • v.37 no.4
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    • pp.354-357
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    • 2021
  • In Korean waters, the cirolanid isopod, Eurydice longiantennata Nunomura and Ikehara, 1985 has been reported only from the subtidal zone of Jeju island. We obtained the mitochondrial cytochrome c oxidase subunit I (COI) sequences of this species and determined the DNA barcoding data of E. longiantennata based on a genetic comparison of E. longiantennata and its congeners. The intra-specific genetic distance between the three COI sequences of E. longiantennata ranged from 0 to 0.6%. The inter-specific distances between E. longiantennata and other cirolanid isopods ranged from 24 to 33.2%. In this study, we provided the DNA information of E. longiantennata with a morphological diagnosis and images of the species.

Peptoniphilus mikwangii-specific quantitative real-time polymerase chain reaction primers

  • Park, Soon-Nang;Kook, Joong-Ki
    • International Journal of Oral Biology
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    • v.44 no.3
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    • pp.96-100
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    • 2019
  • The purpose of this study was to develop Peptoniphilus mikwangii-specific quantitative real-time polymerase chain reaction (qPCR) primers based on the 16S ribosomal RNA (16S rDNA) gene. The specificity of the primers was determined by conventional PCR using 29 strains of 27 oral bacterial species including P. mikwangii. The sensitivity of the primers was determined by qPCR using the purified genomic DNA of P. mikwangii KCOM $1628^T$ (40 ng to 4 fg). The data showed that the qPCR primers (RTB134-F4/RTB134-R4) could detect P. mikwangii strains exclusively and as little as 40 fg of the genomic DNA of P. mikwangii KCOM $1628^T$. These results suggest that the developed qPCR primer pair can be useful for detecting P. mikwangii in epidemiological studies of oral bacterial infectious diseases.

DNA Barcoding of a Colonial Ascidian, Botrylloides violaceus (Ascidiacea: Stolidobrachia: Styelidae), from South Korea

  • Lee, Taekjun;Shin, Sook
    • Animal Systematics, Evolution and Diversity
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    • v.37 no.1
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    • pp.26-30
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    • 2021
  • Botrylloides violaceus is native to the Northwest Pacific, including Korea. This species has many color variations in alive condition and a variety of zooid compound forms, and therefore difficult to identification in the field survey. This is the first report of COI DNA barcodes of B. violaceus from Korea. The intra-specific pairwise distance between Korean and UK populations had ranged from 1.4% to 2.6%. The inter-specific variations between B. violaceus and other Botrylloides species were 21.0-36.8%. The new DNA barcodes for Korean B. violaceus may be helpful in the identification of colonial ascidians, which is a difficult task when based on morphological identification.

Nucleotide Sequence of Rat Transketolase and Liver-Specific Pretranslational Activation During Postnatal Development

  • Kim, Sung-Min F.;Kim, Byung-Moon;Jeng, Jingjau;Soh, Yun-Jo;Bak, Choong-Il;Huh, Jae-Wook;Song, Byoung-J.
    • BMB Reports
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    • v.29 no.2
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    • pp.146-150
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    • 1996
  • A 2.1 kb cDNA clone for rat transketolase was isolated from rat liver ${\lambda}gt11$ cDNA library and its sequence was determined. The predicted rat transketolase (655 amino acids with $M_r$ 71,186) is highly similar (92%) to that of the human enzyme except that it contains an extra 32 amino acids at its N-terminus. Although it is less similar (<27%) to transketolases from non-mammalian species, the functional motifs such as the catalytic sites and thiamine binding domain are well conserved in the rat enzyme. Southern blot analysis of genomic DNA verified that transketolase appears to be derived from a single gene. Immunoblot and Northern blot analyses suggested that hepatic transketolase was activated pretranslationally by a 2.1-fold while little change was observed in brain enzyme, indicating a tissue-specific pretranslational activation during postnatal development.

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A Genetic Marker for the Korean Native Cattle (Hanwoo) Found by an Arbitrarily Primed-Polymerase Chain Reaction (AP-PCR)

  • Lee, Ji-Seon;Lee, Chang-Hee;Nam, Doo-Hyun;Jung, Young-Ja;Yeo, Jung-Sou
    • BMB Reports
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    • v.33 no.3
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    • pp.208-212
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    • 2000
  • In order to develop a specific genetic marker for the Korean native cattle (Hanwoo), an arbitrarily-primed polymerase chain reaction (AP-PCR) analysis of 6 different cattle breeds was attempted. Eight different arbitrary primers, each longer than 20-mer nucleotides, were used. In comparison to the AP-PCR patterns, several distinctive DNA bands that are specific for a certain breed were detected. When the primer Kpn-X was employed, a 280bp DNA fragment was found to be specific only for Hanwoo. In an individual analysis of Hanwoo, this AP-PCR marker was observed in 123 head of cattle among the 153 that were tested (80.4%). Nucleotide sequencing revealed that this fragment has a short microsatellite sequence of tandem repeat, $A(G)_{1-2}\;(C)_{1-3}AGAG$. According to the analysis of AP-PCR band patterns, Hanwoo was discovered to be genetically most closely-related with Holstein among the various cattle breeds.

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