• 제목/요약/키워드: Natural DNA

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작잠(Antheraea pernyi) 아릴포린(Arylphorin) 유전자의 cDNA 클로닝 및 아릴포린 유전자의 발육시기 의존성 발현양상 (cDNA Cloning and Stage-Dependant Expression of Arylphorin Gene from Chinese Oak Silkworm, Antheraea pernyi)

  • 이상몽;황재삼;박남숙;김용균;김근기;손홍주;박현철;진병래
    • 생명과학회지
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    • 제20권8호
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    • pp.1193-1200
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    • 2010
  • 상수리 잎을 먹고 자라는 야생견사곤충의 일종인 작잠의 저장단백질인 아릴포린 유전자의 cDNA를 클로닝하고 발육경과에 따른 유전자발현의 양상을 조사 검토하였다. 작잠의 아릴포린 유전자의 cDNA는 2,112 bp의 ORF(open reading frame)를 포함하여 2,234 bp임을 밝혔다. 작잠 아릴포린 유전자의 cDNA염기서열로부터 아미노산 서열을 검토한 결과 방향족 아미노산인 페닐알라닌(phenylalanine)과 티로신(tyrosine)의 성분이 높은 아미노산 서열구조를 보였으며 ORF로부터 계산한 단백질의 분자량은 83,439 Da 이었다.작잠 아릴포린 저장단백질의 아미노산서열을 다른 곤충의 서열과 그 상동성을 비교 분석한 결과 세크로피아잠(H. cecropia)과는 78%, 담배나방의 알파단량체(M. sexta-$\alpha$ subunit)와는 71%, 담배나방의 베타단량체(M. sexta $\beta$-subunit)와는 62% 그리고 가잠(B. mori)과는 64%의 아미노산서열 상동성을 각각 보였다. 또, Northern blot analysis에서 유충 5령기의 중장, 중부 실샘, 후부 실샘에서는 아릴포린 유전자가 발현되지 않고 오로지 지방체 조직에서만 아릴포린 유전자가 발현되었음을 확인하였다. 아릴포린 유전자는 특히 종령인 5령 유충의 초기에 발현강도가 높았으며 중, 후기로 갈수록 그 강도가 감소하였다. 하지만 번데기시기에는 흔적 정도의 해당 mRNA가 검출되었으며, 이와 같이 검출된 mRNA는 불활성화된 것으로 추정된다. 이상의 결과에서 작잠 아릴포린 유전자의 cDNA가 클로닝되었으며, 이 유전자는 유충기특이적 발현 및 지방체 조직특이적 발현양상을 보인 점이 본 연구에서 확인되었다.

Adjuvant effect of liposome-encapsulated natural phosphodiester CpG-DNA

  • Kim, Dong-Bum;Kwon, Sang-Hoon;Ahn, Chi-Seok;Lee, Young-Hee;Choi, Soo-Young;Park, Jin-Seu;Kwon, Hyeok-Yil;Kwon, Hyung-Joo
    • BMB Reports
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    • 제44권11호
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    • pp.758-763
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    • 2011
  • Immunostimulatory CpG-DNA targeting TLR9 is one of the most extensively evaluated vaccine adjuvants. Previously, we found that a particular form of natural phosphodiester bond CpG-DNA (PO-ODN) encapsulated in a phosphatidyl-${\beta}$-oleoyl-${\gamma}$-palmitoyl ethanolamine (DOPE) : cholesterol hemisuccinate (CHEMS) (1 : 1 ratio) complex (Lipoplex(O)) is a potent adjuvant. Complexes containing peptide and Lipoplex(O) are extremely useful for B cell epitope screening and antibody production without carriers. Here, we showed that IL-12 production was increased in bone marrow derived dendritic cells in a CpG sequence-dependent manner when PO-ODN was encapsulated in Lipoplex(O), DOTAP or lipofectamine. However, the effects of Lipoplex(O) surpassed those of PO-ODN encapsulated in DOTAP or lipofectamine and also other various forms of liposome-encapsulated CpG-DNA in terms of potency for protein antigen-specific IgG production and Th1- associated IgG2a production. Therefore, Lipoplex(O) may have a unique potent immunoadjuvant activity which can be useful for various applications involving protein antigens as well as peptides.

Analysis of the Oxidative Stress-Related Transcriptome from Capsicum annuum L.

  • Lee, Hyoung-Seok;Lee, Sang-Ho;Kim, Ho-Bang;Lee, Nam-Houn;An, Chung-Sun
    • Journal of Plant Biotechnology
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    • 제37권4호
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    • pp.472-482
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    • 2010
  • For the massive screening of the genes related to oxidative stress, a cDNA library was constructed from hot pepper (Capsicum annuum L. cv. Nockkwang) leaves treated with methyl viologen. From this library, 1,589 cDNA clones were sequenced from their 5' ends. The sequences were clustered into 1,252 unigenes comprised of 152 contigs and 1,100 singletons. Similarity search against NCBI protein database identified 1,005 ESTs (80.3%) as Known, 197 ESTs (15.7%) as Unknown, and 50 ESTs (3.99%) as No hit. In the ESTs, oxidative stress-related genes such as ascorbate peroxidase, catalase, and osmotin precursor were highly expressed. The cDNA microarray containing 1,252 unigenes was constructed and used to analyze their expression upon methyl viologen treatment. Analyses of the hybridization revealed that various stress-related genes such as peroxidase, tyrosine aminotransferase, and omega-6 fatty acid desaturase, were induced and some metabolism related genes such as aldolase and ketol-acid reductoisomerase, were repressed by methyl viologen treatment, respectively. The information from this study will be used for further study on the functional roles of oxidative stress-related genes and signaling network of oxidative stress in hot pepper.

Bioluminescence capability and intensity in the dinoflagellate Alexandrium species

  • Park, Sang Ah;Jeong, Hae Jin;Ok, Jin Hee;Kang, Hee Chang;You, Ji Hyun;Eom, Se Hee;Yoo, Yeong Du;Lee, Moo Joon
    • ALGAE
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    • 제36권4호
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    • pp.299-314
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    • 2021
  • Some species in the dinoflagellate genus Alexandrium are bioluminescent. Of the 33 formally described Alexandrium species, the bioluminescence capability of only nine species have been tested, and eight have been reported to be bioluminescent. The present study investigated the bioluminescence capability of seven Alexandrium species that had not been tested. Alexandrium mediterraneum, A. pohangense, and A. tamutum were bioluminescent, but A. andersonii, A. hiranoi, A. insuetum, and A. pseudogonyaulax were not. We also measured the bioluminescent intensity of A. affine, A. fraterculus, A. mediterraneum, A. ostenfeldii, A. pacificum, A. pohangense, A. tamarense, and A. tamutum. The mean 200-second-integrated bioluminescence intensity per cell ranged from 0.02 to 32.2 × 104 relative luminescence unit per cell (RLU cell-1), and the mean maximum bioluminescence intensity per cell per second (BLMax) ranged from 0.01 to 10.3 × 104 RLU cell-1 s-1. BLMax was significantly correlated with the maximum growth rates of Alexandrium species, except for A. tamarense. A phylogenetic tree based on large subunit ribosomal DNA (LSU rDNA) showed that the bioluminescent species A. affine, A. catenella, A. fraterculus, A. mediterraneum, A. pacificum, and A. tamarense formed a large clade. However, the toxicity or mixotrophic capability of these species was split. Thus, their bioluminescence capability in this clade was more consistent than their toxicity or mixotrophic capability. Phylogenetic trees based on LSU rDNA and the luciferase gene of Alexandrium were consistent except for A. pohangense. The results of the present study can provide a basis for understanding the interspecific diversity in bioluminescence of Alexandrium.

Collaborative Effect of CuZnSOD and Human AP Endonuclease against Oxidative Stress

  • Kim Young Gon
    • 한국미생물학회:학술대회논문집
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    • 한국미생물학회 2004년도 International Meeting of the Microbiological Society of Korea
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    • pp.47-50
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    • 2004
  • The defenses against free radical damage include specialized repair enzymes that correct oxidative damages in DNA, and detoxification systems such as superoxide dismutases. These defenses may be coordinated genetically as global responses. We hypothesized that the expression of the SOD and the DNA repair genes would inhibit DNA damage under oxidative stress. Therefore, the protection of E. coli mutants deficient in SOD and DNA repair genes $(sod^-\;xth^-\;and\;nfo^-)$ was demonstrated by transforming the mutant strain with a plasmid pYK9 which encoded Photobacterium leiognathi CuZnSOD and human AP endonuclease. The results show that survival rates were increased in $sod^+\;xth^-\;nfo^+$ cells compared to $sod^-\;xth^-\;ap^+,\;sod^-\;xth^-\;ap^-,\;and\;sod^+\;xth^-\;ap^-$ cells under oxidative stress generated from 0.1 mM Paraquat or 3 mM $H_2O_2$. The data suggested that, at least, SOD and DNA repair enzymes may have collaborate protection and repair of the damaged DNA. Additionally, both enzymes are required for protection against free radicals.

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