• 제목/요약/키워드: CytA

검색결과 109건 처리시간 0.024초

Amino acids at N- and C-termini are required for the efficient production and folding of a cytolytic γ-endotoxin from Bacillus thuringiensis

  • Thammachat, Siriya;Pathaichindachote, Wanwarang;Krittanai, Chartchai;Promdonkoy, Boonhiang
    • BMB Reports
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    • 제41권11호
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    • pp.820-825
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    • 2008
  • Bacillus thuringiensis Cyt2Aa toxin is a mosquito-larvicidal and cytolytic $\delta$-endotoxin, which is synthesized as a protoxin and forms crystalline inclusions within the cell. These inclusions are solubilized under alkaline conditions and are activated by proteases within the larval gut. In order to assess the functions of the N-and C-terminal regions of the protoxin, several N- and C-terminal truncated forms of Cyt2Aa were constructed. It was determined that amino acid removal at the N-terminal, which disrupts the $\beta$1 structure, might critically influence toxin production and inclusion formation. The deletion of 22 amino acids from the C-terminus reduced the production and solubility of the toxin. However, the removal of more than 22 amino acids from the C-terminus or the addition of a bulky group to this region could result in the inability of the protein to adopt the proper folding. These findings directly demonstrated the critical roles of N- and C-terminal amino acids on the production and folding of the B. thuringiensis cytolytic $\delta$-endotoxin.

A Phylogenetic Study of Korean Rodents (Muridae, Sciuridae) Based on Mitochondrial and Nuclear DNA

  • Jung, Gi-La;Lee, Seo-Jin;Kim, Chuel-Kyu;Lee, Hang;Kim, Chang-Bae
    • Animal Systematics, Evolution and Diversity
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    • 제26권2호
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    • pp.99-104
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    • 2010
  • The subfamily Murinae is a very controversial group concerning their phylogenetic relationship. Previous studies could not resolve phylogeny among four genera Apodemus, Micromys, Mus and Rattus of the Muridae. In the present study, eight rodent species resident in South Korea were collected and phylogenetically analyzed based on sequence data of five mitochondrial and nuclear DNA regions: 12S rRNA, cytochrome b gene (cyt b), cytochrome oxidase II (COII), control region of mitochondrial DNA, and a thyroglobulin (Tg) of nuclear DNA. According to the phylogeny of the concatenated data, M. musculus separated early in Murinae (ML 100%; BA 1.00 pp) and the genus Rattus grouped with the harvest mouse, M. minutes; these were separated from the genus Apodemus with relatively strong support (ML 74%; BA 0.76 pp). The Siberian chipmunk population was also examined using the five genes to obtain better resolution. The phylogeny for Korean rodents determined using the 12S rRNA, cyt b, COII and control regions discriminated the Siberian chipmunk populations from Korea, Russia, and China.

Expression and Purification of GFPuv/Cytochrome c-552 Fusion Protein in E. coli

  • 홍을재;이상온;최정우;홍억기
    • 한국생물공학회:학술대회논문집
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    • 한국생물공학회 2003년도 생물공학의 동향(XII)
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    • pp.550-553
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    • 2003
  • $Hisx_6-GFPuv-Cyt$ c fusion protein을 E. coli 균주 JM109과 BL21 각각에서 발현시킨 결과 발현온도는 $30^{\circ}C$보다 $37^{\circ}C$에서, BL21보다 JM109에서의 발현량이 더 많았다. 그러나 JM109, $37^{\circ}C$에서 발현시킨 fusion protein의 $Ni^{2+}-IDA-agarose$ purification결과 약 45kDa 부근의 fusion protein의 density가 감소되었음을 SDS-PAGE analysis을 통해 알 수 있었다. 또한 western blotting analysis를 통해 이 impurity가 degraded fusion임을 확인 할 수 있었다. degraded fusion은 BL21 균주에서 발현시킨 fusion protein에서도 생성됨을 확인하였다. 모든 결과를 종합해 볼때 $Hisx_6-GFPuv-Cyt$ c fusion protein의 발현은 IM109, $37^{\circ}C$에서 더 많았지만, BL21, $37^{\circ}C$에서 expression시킨 fusion protein이 보다 안정하다고 판단 되어진다. 향후 fusion protein이 bioelectronic device에 적용되려면 degraded fusion protein의 생성을 줄여 activity를 유지하도록 안정한 형태로 발현되어 순수하게 분리정제 되어야 하겠다.

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Molecular Phylogeny of the Gayal in Yunnan China Inferred from the Analysis of Cytochrome b Gene Entire Sequences

  • Li, S.P.;Chang, H.;Ma, G.L.;Cheng, H.Y.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권6호
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    • pp.789-793
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    • 2008
  • The gayal (Bos frontalis) in China is a very rare semi-wild and semi-domestic bovine species. There still exist remarkable divergences on the gayal's origin and taxonomic status. In the present study, the cytochrome b (Cyt b) gene entire sequences (1,140 bp) of 11 gayals in Yunnan China were analyzed. Combined with other bovine Cyt b sequences cited in GenBank, the phylogenetic trees of genus Bos were reconstructed by neighbor-joining (NJ) and maximum parsimony (MP) methods with Bubalus bubalis as outgroup. Sequence analysis showed that, among 1,140 sites compared for 11 gayals, 95 variable sites (8.33% of all sites) and 6 different haplotypes were observed, showing abundant mitochondrial genetic diversity in gayals. Both NJ and MP trees demonstrated that gayals in this study were markedly divided into three embranchments: one embranchment clustering with Bos gaurus, another clustering with Bos taurus, and the third clustering with Bos indicus. The result of phylogenetic analysis suggested that the gayal might be the domesticated form of the gaur, and a great proportion of the gayal bloodline in China was invaded by other bovine species.

담배 (Nicotiana tabacum L.) 현탁배양세포의 Elicitor 유도성 5-epi-Aristolochene Hydroxylase (Elicitor-Inducible 5-epi-Aristolochene Hydroxylase in Suspension Cultures of Tobacco (Nicotiana tabacum L.))

  • KWON, Soon-Tae;CHAPPELL, Joseph
    • 식물조직배양학회지
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    • 제25권3호
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    • pp.141-146
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    • 1998
  • [$^3$H] 5-epi-aristolochene (5-EAS)를 담배 현탁배양세포에 투여하여 elicitor에 의해 유도된 세포가 생합성하여 배지로 방출하는 [$^3$H]-capsidiol의 량을 측정함으로써 5-EAS hydroxylase의 활성을 검정하였고, 이 반응의 전 단계에 관여하는 효소인 sesquiterpene cyclase의 발현특성과 비교하였다. 5-EAS hydroxylase는 정상세포에는 전혀 활성을 보이지 않으나 elicitor로써 cellulase를 처리한 세포는 9시간 후부터 유도를 시작하여 18시간 후에 최대 활성을 보였는데 동일한 세포내에서 유도되는 cyclase와 유사한 패턴을 보였다. Cyt P450계 효소의 특이적 억제제로 알려진 ancymidol과 ketoconazole에 의해 5-EAS hydroxylase의 활성은 강한 억제를 보인 반면 cyclase의 활성은 억제를 보이지 않아 5-EAS hydroxylase가 P450계 효소임이 시사되었다.

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참복속(genus Takifugu) 어류 3종과 미동정 1종의 형태 및 유전학적 비교 (Morphological and Molecular Comparison among Three Species and One Unidentified Takifugu Species)

  • 백정익;한경호;이성훈;김진구
    • 한국수산과학회지
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    • 제51권4호
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    • pp.404-410
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    • 2018
  • Takifugu rubripes, T. chinensis, T. pseudommus and an unidentified species were morphologically and genetically studied. Morphology was compared using external morphology and the genetic analysis was conducted using mitochondrial DNA cytochrome oxidase I (COI) and cytochrome b (Cyt b). The morphological characteristics of T. rubripes, T. chinensis and T. pseudommus were confirmed, as reported previously. The unidentified species was confirmed to have the characteristics of T. rubripes and T. chinensis. The COI sequences of the four species were 99-100% similar; genetic distance was d=0.0000 and Cyt b genetic distance was d=0.0000-0.00834. Species in the phylogenetic tree belonging to the same group could not be classified genetically. In conclusion, the unidentified species was considered to be a hybrid between T. rubripes and T. chinensis.

Effects of Psidium guajava Leaf Extract on Apoptosis Induction Through Mitochondrial Dysfunction in HepG2 Cells

  • Nguyen, Van-Tinh;Ko, Seok-Chun;Oh, Gun-Woo;Heo, Seong-Yeong;Jung, Won-Kyo
    • 한국미생물·생명공학회지
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    • 제47권1호
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    • pp.43-53
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    • 2019
  • The anticancer activity of guava (Psidium guajava L.) leaf extract (GLE) occurs via the induction of apoptosis in cancer cells. However, the mechanism behind GLE-induced apoptosis in the human hepatocellular carcinoma cell line HepG2 remains unclear. In the present study, we investigated the apoptotic effects and mechanism of action of GLE in cultured HepG2 cells. The results showed that GLE induced reactive oxygen species (ROS) synthesis and disrupted the mitochondrial membrane potential (${\Delta}{\Psi}m$). Moreover, GLE increased the expression of apoptotic pathway proteins, such as the cleaved forms of caspase-3, -8, and -9; the translocation of Bax and cytochrome c (cyt-c) from the mitochondria to the cytosol; and the downregulation of Bcl-2. In addition, p53 protein expression was increased upon GLE treatment. These observations indicate that the GLE-induced apoptosis in HepG2 cells is mediated by mitochondrial ROS generation, followed by caspase activation and cyt-c release, suggesting that GLE may be a promising candidate for the development of novel drugs for the treatment of liver cancers.

녹색자엽 검정콩 품종 육성을 위한 Stay green 변이체 활용 (Introduction of Stay Green Mutant for the Development of Black Seed Coat and Green Cotyledon Soybean Variety)

  • 강성택;서민정;문중경;윤홍태;이영호;김시주;황영선;이석기;정명근
    • 한국작물학회지
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    • 제55권3호
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    • pp.187-194
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    • 2010
  • 고품질 녹색자엽 검정콩 신품종 육성 연구의 일환으로 녹색자엽의 유전자원별 녹색의 정도를 평가할 수 있는 엽록소 함량 및 조성과 이들의 유전양상을 검토하고, 유전적 배경을 달리한 녹색자엽 자원을 활용한 녹색자엽 검정콩의 선발 효율을 검토한 결과를 요약하면 아래와 같다. 1. 유전적 배경이 상이한 녹색자엽 자원을 검정콩 육종프로그램에 도입코자 실시한 특성조사 결과 핵유전 녹색자엽 유전자 d1d2를 보유한 자원은 세포질 유전 녹색자엽 유전자 cytG를 보유한 자원보다 총 엽록소의 함량이 매우 높은 경향이었고, 조성에서는 d1d2 보유 자원은 엽록소 a의 함량이 엽록소 b보다 2배 이상 높고, cytG 보유 유전자원은 엽록소 b의 함량이 엽록소 a 함량 보다 높은 경향이었다. 2. 핵유전 녹색자엽 유전자(d1d2)는 당대의 종실에서 발현되어 $F_1$ 세대의 $F_2$ 종실에서부터 선발이 가능하며, 2개의 열성유전자에 지배를 받는다. 3. 녹색 종피의 핵유전 녹색자엽(d1d2)과 황색자엽 검정콩(D1D2)의 교배후대에서 검정색 종피에 녹색자엽 종자를 가진 개체는 모본의 유전적 배경에 따라 조금은 다르지만 $F_2$ 전체 개체 중 3% 미만의 낮은 비율로 나타나, 향후 핵유전 녹색자엽 검정콩 신품종 육성효율 증진을 위해서는 핵유전 녹색자엽 검정콩 중간모본 개발과 선발방법 개선이 필요할 것으로 판단된다.

DNA와 Benzo(a)pyrene 대사물질 결합형성에 미치는 인삼 추출물의 영향 (Effect of Ginseng Extracts on the Binding to DNA of Benzo(a)pyrene Metabolites in uitro in Rats)

  • 박진규;고지훈
    • Journal of Ginseng Research
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    • 제13권1호
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    • pp.37-41
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    • 1989
  • Benzo(a)pyrene(BP)의 monooxygenase(AHH)에 의해서 생성된 반응성 대사물질들의 in vitro DNA와의 결합 및 BP 대사에 관여하는 효소들의 활성도에 미치는 인삼 물추출물의 영향을 조사하였으며, DNA-BP metabolite adduct들은 Sephadex LH-20 column으로 chromatography하여 5개의 major peak 들을 얻었다. 이 peak 들을 극성이 큰 순서대로 A부터 E까지 임의로 정하고 5개의 peak들을 7,8-diol-9,10-oxide(A), 7,8-oxide(B). 4,5-oxide(C), 9-HO-BP(D & E) adduct들로 잠정적으로 확인하였다. Peak A, C, D 그리고 E는 각각 대조군의 30, 15, 20 그리고 30%로 감소되었으며 peak B는 의미있는 변화를 보이지 않았다. DNA-BP 결합 억제와 관련하여 in vitro와 in vivo 투여시의 경향이 유사하여 EH의 활성도만 BP투여 대조군보다 38%정도 의미있게 유도되었다.

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The Expression of a Cytosolic Fructose-1,6-Bisphosphatase, a Key Enzyme in Sucrose Biosynthesis, Gene was Diurnally Fluctuated and Increased in Cold Acclimated Leaves of Chinese Cabbage

  • Leen, Jeong-Yeo;Song, Ha-Young;Lim, Yong-Pyo;Hur, Yoon-Kang
    • Journal of Plant Biotechnology
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    • 제33권2호
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    • pp.123-131
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    • 2006
  • Chinese cabbage (Brassica rapa ssp. pekinesis) is one of the most important vegetable crops in korea and other East Asian countries. Cytosolic fructose-1,6-bisphospha-tase (cytFBPase) is a key enzyme in sucrose biosyn-thesis, which controls the sucrose levels as well as the productivity at plants. The Chinese cabbage cytFBPase gene, BrFBPase, encodes the 340 amino acid polypep-tide, giving a theoretical molecular weight of 37.2 kD and a isolectric point of 5.4. BrFBPase showed high sequence identity with Brassica homologs and its functional domains, such as 12,6P$_2$ binding site or active site and F6P binding site, were highly conserved in diverse sources of organisms. Although the genome of Chinese cabbage seemed to be triplicated, BrFBPase appears to be a single copy gene. The expression of BrFBPase was examined at transcript and protein levels under various conditions. BrFBPase expression was observed only in photosynthetic source tissue, not in sink tissue. The expression was slightly higher during the day than at night, and it showed a diurnal cycle with circadian rhythmicity. Short-term exposure to low temperature inhibited the expression of the BrFBPase, while long-term exposure increased the expression, supporting that sugar levels are high in late autumn when temperature are low.