• 한국약용작물학회지
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• 제13권5호
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• pp.270-275
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• 2005

Superoxide dismutases (SOD) are metalloenzymes that convert $O_2^-\;to\;H_2O_2$. Rehmannia glutinosa is highly tolerant to paraquat-induced oxidative stress. The primary objective of this study was to characterize regulation of SOD gene expression in R. glutinosa in response to oxidative stresses and hormones. A full-length putative SOD clone (RgCu-ZnSOD1) was isolated from the leaf cDNA library of R. glutinosa using an expressed sequence tag clone as a probe. RgCu-ZnSOD1 cDNA is 777 bp in length and contains an open reading frame for a polypeptide consisted of 152 amino acid residues. The deduced amino acid sequence of the clone shows highest sequence similarity to the cytosolic Cu-ZnSODs. The two to three major bands with several minor ones on the Southern blots indicate that RgCu-ZnSOD1 is a member of a small multi-gene family. RgCuZnSOD1 mRNA was constitutively expressed in the leaf, flower and root. The expression of RgCu-ZnSOD1 mRNA was increased about 20% by wounding and paraquat, but decreased over 50% by ethylene and $GA_3$. This result indicates that the RgCu-ZnSOD1 expression is regulated differentially by different stresses and phytohormones at the transcription level. The RgCu-ZnSOD1 sequence and information on its regulation will be useful in investigating the role of SOD in the paraquat tolerance of R. glutinosa.

• International Journal of Industrial Entomology and Biomaterials
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• 제5권1호
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• pp.85-91
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• 2002

The sequences of the internal transcribed spacers (ITS1 and ITS2) and 5.8S ribosomal DNA gene from five Cordyceps species and one Paecilomyces japonica were determined. The total length of the ITSI, 5.8S and ITS2 regions ranged from 528 to 549 bp. When the C. militaris collected from Korea was used as a standard genotype, the sequence showed 88.4%, 88.6%, 91.1% and 86.8% identity to C. pruinosa, C. sphecocephala, C. scarabaeucika and R japonica, respectively, while the lowest identity was found with C. sinensis (75.4%). Interestingly, C. sinensis was phylogenetically distant from the other Cordyceps species. To test geographic variation, furthermore, sequences of the ITS regions in the 8 samples of C. militaris collected from two localities in Korea and China analyzed and compared with the GenBank-searched sequences from Japan and China. The total length of the ITS regions of C. militaris from Korea, Japan and China was completely identical to each other with 528 bp, and the sequence divergence among three localities in pairwise comparisons ranged from 0.2% (1 bp) to 0.4% (2 bp).

• 한국미생물·생명공학회지
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• 제31권2호
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• pp.117-123
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• 2003

공시균인 Saccha. erythraea IFO 13426으로부터 VB-C에 의한 erythromycin 생산 유도능이 시사된 바 있으므로, 공시균으로부터 VB-C와 특이적으로 결합하는 autoregulators 및 receptor gene을 탐색하여, EM의 생산 조절 기구를 규명하고자 하였다. 탐색의 일환으로 기존의 Streptomyce속 receptor gene의 공통배열을 primer로 이용하여 PCR을 수행하였고, 예상 크기인 120bp의 단편을 pUC19 vector에 ligation하여 E. coli DH5$\alpha$에 형질전환한 후, plasmid를 분리하여 BamHI을 처리하여 2% agarose gel에 전기영동한 결과, pUC19 (2.7kbp)외에 receptor gene PCR 산물이 120bp위치에 존재하는 것을 확인하였다. 형질전환된 plasmid로 PCR을 수행하여 염기배열을 결정한 후 해석한 결과 Streptomyces sp. 유래의 receptor gene과 유사함을 확인하였다. 따라서 Saccha. erythraea IFO 13426에는 항생물질인 erythromycin의 생산에 관여한다고 추정되는 autoregulator receptor protein을 코드하는 유전자가 존재할 것으로 예상되어 120 bp의 PCR product를 probe로 이용하여 Southern 및 colony hybridization을 통하여 3.2 kbp의 SacI 단편을 가지는 plasmid(pESG)를 제작하였고, 이를 sequencing한 결과, autoregulator receptor protein 유전자가 KpnI과 SalI을 포함하는 영역에 존재한다는 것을 알 수 있었으며 이를 EsgR이라 명명하였다. 유전자 해석 결과, EsgR은 205개의 아미노산으로 구성되어 있으며, 이는 기존의 autoregulator receptor proteins과 비교시 30%이상의 상동성을 나타내었으며, 기존의 autoregulator receptor prorein들이 하부의 항생물질 생합성 유전자들의 제어를 위해 보유하고 있는 helix-turn-helix DNA binding motif를 EsgR이 보유하고 있는 점에서, EsgR은 Saccha. erythraea가 보유하는 autoregulator receptor protein을 code하는 유전자로 추정되었다.

• Journal of Microbiology and Biotechnology
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• 제14권2호
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• pp.276-283
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• 2004

Screening was performed to isolate cellulose-producing microorganisms from the Korean traditional fermented persimmon vinegar. The resulting strain, KJ $145^{T}$, was then taxonomically investigated by phenotypic characterization, particularly chemotaxonomic, and by phylogenetic inference based on a 16S rDNA sequence analysis including other related taxa. Strain KJ $145^{T}$ was found to grow rapidly and form pale white colonies with smooth to rough surfaces on a GYC agar. Strain KJ $145^T$ also produced acetate from ethanol, and was tolerable to 10% ethanol in SM medium. In a static culture, a thick cellulose pellicle was produced, and in GYC broth, the strain grew at temperatures ranging from 28 to $40^\circ{C}$ with an optimum pH of 4.0. The genomic DNA G+C content of strain KJ $145^T$ was 61.9 mol%, and the predominant ubiquinone was Q 10 as the major quinone and Q9 as the minor quinone. The major cellular fatty acids were $C_{16:0}$ and the sum in feature 7 ($C_{18:1}$ w9c, w12t and/or w7c). A 16S rRNA-targeted oligonucleotide probe specific for strain KJ $145^T$was constructed, and the phylogenetic position of the new species was derived from a 16S rDNA-based tree. When comparing the 16S rDNA nucleotide sequences, strain KJ $145^T$ was found to be most closely related to G. hansenii LMG $1527^T$ (99.2%), although KJ $145^T$ was still distinct from G. hansenii LMG $l527^T$ and G. xylinus LMG $1515^T$ in certain phenotypic characteristics. Therefore, on the basis of 16S rDNA sequences and taxonomic characteristics, it is proposed that strain KJ $145^T$ should be placed in the genus Gluconacetobacter as a new species, Gluconacetobacter persimmonis sp. nov., under the type-strain KJ $145^T$ (=KCTC =$10175BP^T$=KCCM=$10354^T$).

• 대한본초학회지
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• 제30권3호
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• pp.41-47
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• 2015

Objectives : Due to the morphological similarity of the pericarp and description of multi-species in National Pharmacopoeia of Korea and China, the Zanthoxylum Pericarpium is difficult to authenticate adulterant in species levels. Therefore, we introduced the sequence analysis of DNA barcode and identification of single nucleotide polymorphism(SNP) to establish a reliable tool for the distinction of Zanthoxylum Pericarpium from its adulterants. Methods : To analyze DNA barcode region, genomic DNA was extracted from twenty-four specimens of authentic Zanthoxylum species and inauthentic adulterant and the individual internal transcribed spacer regions (rDNA-ITS and ITS2) of nuclear ribosomal RNA gene were amplified using ITS1, ITS2-S2F, and ITS4 primer. For identification of species-specific sequences, a comparative analysis was performed using entire DNA barcode sequences. Results : In comparison of four Zanthoxylum ITS2 sequences, we identified 16, 4, 6, and 4 distinct species-specific nucleotides enough to distinguish Z. schinifolium, Z. bungeanum, Z. piperitum, and Z. simulans, respectively. The sequence differences were available genetic marker to discriminate four species. Futhermore, phylogenetic relationship revealed a clear classification between different Zanthoxylum species showing 4 different clusters. These results indicated that comparative analysis of ITS2 DNA barcode was an useful genetic marker to authenticate Zanthoxylum Pericarpium in species levels. Conclusions : The marker nucleotides, enough to distinguish Z. schinifolium, Z. piperitum, Z. bungeanum, and Z. simulans, were obtained at 30 SNP marker nucleotides from ITS2 sequences. These differences could be used to authenticate official Zanthoxylum Pericarpium from its adulterants as well as discriminating each four species.

• Animal Systematics, Evolution and Diversity
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• 제31권3호
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• pp.182-190
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• 2015

Two heterotrichid ciliates, Climacostomum virens (Ehrenberg, 1838) Stein, 1859 from brackish water and freshwater, and Fabrea salina Henneguy, 1890 from a solar saltern, were collected in Korea. They are novelly investigated in Korea by means of live observation, protargol staining and nuclear small subunit (SSU) rRNA gene sequencing. Climacostomum virens is characterized by pouch-like body shape, body length of $200-370{\mu}m$ in vivo, conspicuous cytopharyngeal tube, macronuclei ribbon-like shape, and one to four in number, with or without symbiont algae in cytoplasm, 34-66 somatic kineties, 67-113 adoral zone of membranelles, 8-42 peristomial kineties, 24-37 apical membranelles. SSU rDNA sequence size is 1,591 bp and GC contents 48.52%. Fabrea salina is also characterized by scoop-like body shape with proboscis, body length of $190-240{\mu}m$ in vivo, one to two rod-shaped macronuclei, oval micronuclei, grayish green cortical granules, 104-186 somatic kineties, 4-8 preoral kineties, 7-19 peristomial kineties and fragmented paroral membrane. SSU rDNA sequence size is 1,598 bp and GC contents 47.50%.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.18-24
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• 2003

벼과식물과 관련된 질소고정균을 분리하기 위하여 한반도 남부지방의 19지역에서 쌀, 밀, 귀리, 보리, 호밀, 옥수수 등을 채집하여 이들의 뿌리로부터 free living bacteria와 endophyte를 분류하였다. Colony의 형태, 크기, 색깔 등과 16S rDNA 염기서열을 근거로 63종으로 분류하였다. nifH 유전자 염기서열 결정과 nitrogenase 활성 측정을 통하여 분리균주 모두 질소고정 능력이 있음을 확인하였다. 그리고 식물에서 분리한 내생균의 특징 중 하나인 cellulase활성이 대부분의 분리 균주에서 확인되었다.

• Journal of Microbiology
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• 제37권4호
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• pp.193-199
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• 1999

Fifty-eight strains showing different colony morphological characteristics on various media were isolated from marine coral (Plexauridae sp.) and ambient seawater near Mun-Sum, Cheju-Island in 1998. Bacterial diversity was studies by phylogenetic analysis of the partial 16S rRNA gene sequences. All isolates representing the bacterial domain included affiliates of the high G+C (59%) and los G+C (3%) subdivision of Gram positive bacteria, and the alpha (33%) and gamma (5%) subdivision of the Proteobacteria. The 16S rDNA sequence similarity of the isolates was in the 88.3 to 100% range (average, 95.6%) to reported sequence data. In the comparison of the isolates from marine coarl and ambient seawater, more diverse groups belonging to ${\alpha}$-Proteobacteria were preferentially obtained from seawater.

• 대한수의학회지
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• 제39권4호
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• pp.797-805
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• 1999

T sergenti cDNA library were constructed to get a more broad information about the structural, functional or antigenic properties of the proteins, and analyzes for their partial cDNA sequences and expression sequences tags(ESTg). The mRNA were purified from T sergenti isolates to identify the information of antigen gene, then first and second strand cDNA was synthesized. EcoR I adaptor ligation and Xho I enzyme restriction were used to the synthesized cDNA, and ligated into a Uni-ZAP XR vector. T sergenti cDNA library was constructed with packaging and amplification in vitro. Antibody screening was performed with constructed T sergenti cDNA library using antisera against T sergenti. Among those clones, eight phagemids were rescued from the recombinant in vivo excision with f1 helper phage. Using the analysis of endonuclease restriction and PCR, the recombinant cDNA were proved having a 0.5-3.0kb of inserts. The eight of partial cDNA clones' sequences were obtained and examined for their homology using BLASTN and BLASTX. The eight of sequenced clones were classified into three groups according to the basis of database searches. A total 3,045bp of partial cDNA sequence were determined from six clones. The putatively identified clones contain a cytochrome c gene, a heat shock protein gene, a cyclophilin gene, and a ribosomal protein gene. The unidentified clones have a homology to ATP-binding protein(mtrA) gene of S argillaceus, DNA-binding protein(DBP) gene of Pseudorabies virus 85kDa merozoite protein gene of B bovis, mRNA spm1 protein of T annulata and glycine-rich RNA-binding protein mRNA of O sativa etc.

• 대한의생명과학회지
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• 제6권1호
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• pp.65-72
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• 2000

반추수에서 발생하는 Johne병의 조기 진단 방법을 제시하고 이 질병의 원인체와 미생물학적 특징 이 유사한 M. bovis, M. avium 등의 mycobacteria 감염증을 감별 진단하는 방법 을 개발하기 위하여 Mycobacterium 균속의 표준균주를 사용하여 중합효소 연쇄반응을 확립하였다. Johne병으로 의심되는 소의 혈액과 유즙을 채취하여 분리한 단핵구 및 거식세포로부터 genomic DNA를 추출하였다. 각 시료로부터 추출한 DNA를 template로 이용하여 Mycobacterium spp.에 특이적인 16S rDNA primer set를 이용한 PCR을 수행하여 시료내의 mycobacterial DNA 보유 여부를 확인하였다. 한편 mycobacteria 양성으로 확인된 시료는 M. avium complex 균종에 특이한 16S rDNA 염기서열을 기초로 하여 제작한 primer set와 M. paratuberculosis의 IS900 sequence에 특이한 primer set를 이용하여 duplex PCR을 수행하여 Johne병 원인체의 보균 여부를 조사하였으며, 이 결과를 oligonucleotide probe를 사용한 Southern blot hybridization을 통하여 다시 확인하였다. 이와 같은 duplex PCR기법을 실제 축산 현장에서 수집한 유즙과 말초혈액으로부터 분리한 단핵구 및 거식세포 시료에 적용한 결과 본 연구에서 확립한 duplex PCR기법 유용성을 확인할 수 있었다.